Dual-acting benzoimidazole antihypertensive agents

ABSTRACT

The invention relates to compounds having the formula: 
     
       
         
         
             
             
         
       
     
     wherein Ar, r, n, X, R 2 , R 2′ , R 3 , and R 5-7  are as defined in the specification, or a pharmaceutically acceptable salt thereof. These compounds have AT 1  receptor antagonist activity and neprilysin inhibition activity. The invention also relates to pharmaceutical compositions comprising such compounds; methods of using such compounds; and a process and intermediates for preparing such compounds.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application claims the benefit of U.S. Provisional Application No.61/007,129, filed on Dec. 11, 2007; the disclosure of which isincorporated herein by reference in its entirety.

BACKGROUND OF THE INVENTION

1. Field of the Invention

The present invention relates to novel compounds having angiotensin IItype 1 (AT₁) receptor antagonist activity and neprilysin-inhibitionactivity. The invention also relates to pharmaceutical compositionscomprising such compounds, processes and intermediates for preparingsuch compounds and methods of using such compounds to treat diseasessuch as hypertension.

2. State of the Art

The aim of antihypertensive therapy is to lower blood pressure andprevent hypertension-related complications such as myocardialinfarction, stroke, and renal disease. For patients with uncomplicatedhypertension (i.e., no risk factors, target organ damage, orcardiovascular disease), it is hoped that reducing blood pressure willprevent development of cardiovascular and renal comorbidities,conditions that exist at the same time as the primary condition in thesame patient. For those patients with existing risk factors orcomorbidities, the therapeutic target is the slowing of comorbid diseaseprogression and reduced mortality.

Physicians generally prescribe pharmacological therapies for patientswhose blood pressure cannot be adequately controlled by dietary and/orlifestyle modifications. Commonly used therapeutic classes act topromote diuresis, adrenergic inhibition, or vasodilation. A combinationof drugs is often prescribed, depending upon what comorbidities arepresent.

There are five common drug classes used to treat hypertension:diuretics, which include thiazide and thiazide-like diuretics such ashydrochlorothiazide, loop diuretics such as furosemide, andpotassium-sparing diuretics such as triamterene; β₁ adrenergic receptorblockers such as metoprolol succinate and carvedilol; calcium channelblockers such as amlodipine; angiotensin-converting enzyme (ACE)inhibitors such as captopril, benazepril, enalapril, enalaprilat,lisinopril, quinapril, and ramipril; and AT₁ receptor antagonists, alsoknown as angiotensin II type 1 receptor blockers (ARBs), such ascandesartan cilexetil, eprosartan, irbesartan, losartan, olmesartanmedoxomil, telmisartan, and valsartan. Combinations of these drugs arealso administered, for example, a calcium channel blocker (amlodipine)and an ACE inhibitor (benazepril), or a diuretic (hydrochlorothiazide)and an ACE inhibitor (enalapril). All of these drugs, when usedappropriately, are effective in the treatment of hypertension.Nevertheless, both efficacy and tolerability should be further improvedin new drugs targeting hypertension. Despite the availability of manytreatment options, the recent National Health And Nutrition ExaminationSurvey (NHANES) demonstrated that only about 50% of all treated patientswith hypertension achieve adequate blood pressure control. Furthermore,poor patient compliance due to tolerability issues with availabletreatments further reduces treatment success.

In addition, each of the major classes of antihypertensive agents havesome drawbacks. Diuretics can adversely affect lipid and glucosemetabolism, and are associated with other side effects, includingorthostatic hypotension, hypokalemia, and hyperuricemia. Beta blockerscan cause fatigue, insomnia, and impotence; and some beta blockers canalso cause reduced cardiac output and bradycardia, which may beundesirable in some patient groups. Calcium channel blockers are widelyused but it is debatable as to how effectively these drugs reduce fataland nonfatal cardiac events relative to other drug classes. ACEinhibitors can cause coughing, and rarer side effects include rash,angioedema, hyperkalemia, and functional renal failure. AT₁ receptorantagonists are equally effective as ACE inhibitors but without the highprevalence of cough.

Neprilysin (neutral endopeptidase, EC 3.4.24.11) (NEP), is anendothelial membrane bound Zn²⁺ metallopeptidase found in many tissues,including the brain, kidney, lungs, gastrointestinal tract, heart, andperipheral vasculature. NEP is responsible for the degradation andinactivation of a number of vasoactive peptides, such as circulatingbradykinin and angiotensin peptides, as well as the natriureticpeptides, the latter of which have several effects includingvasodilation and diuresis. Thus, NEP plays an important role in bloodpressure homeostasis. NEP inhibitors have been studied as potentialtherapeutics, and include thiorphan, candoxatril, and candoxatrilat. Inaddition, compounds have also been designed that inhibit both NEP andACE, and include omapatrilat, gempatrilat, and sampatrilat. Referred toas vasopeptidase inhibitors, this class of compounds are described inRobl et al (1999) Exp. Opin. Ther. Patents 9(12): 1665-1677.

There may be an opportunity to increase anti-hypertensive efficacy whencombining AT₁ receptor antagonism and NEP inhibition, as evidenced byAT₁ receptor antagonist/NEP inhibitor combinations described in WO9213564 to Darrow et al. (Schering Corporation); US20030144215 toKsander et al.; Pu et al., Abstract presented at the CanadianCardiovascular Congress (October 2004); and Gardiner et al. (2006) JPET319:340-348; and WO 2007/045663 (Novartis AG) to Glasspool et al.

Recently, WO 2007/056546 (Novartis AG) to Feng et al. has describedcomplexes of an AT₁ receptor antagonist and a NEP inhibitor, where anAT₁ receptor antagonist compound is non-covalently bound to a NEPinhibitor compound, or where the antagonist compound is linked to theinhibitor compound by a cation.

In spite of the advances in the art, there remains a need for a highlyefficacious monotherapy with multiple mechanisms of action leading tolevels of blood pressure control that can currently only be achievedwith combination therapy. Thus, although various hypertensive agents areknown, and administered in various combinations, it would be highlydesirable to provide compounds having both AT₁ receptor antagonistactivity and NEP inhibition activity in the same molecule. Compoundspossessing both of these activities are expected to be particularlyuseful as therapeutic agents since they would exhibit antihypertensiveactivity through two independent modes of action while having singlemolecule pharmacokinetics.

In addition, such dual-acting compounds are also expected to haveutility to treat a variety of other diseases that can be treated byantagonizing the AT₁ receptor and/or inhibiting the NEP enzyme.

SUMMARY OF THE INVENTION

The present invention provides novel compounds that have been found topossess AT₁ receptor antagonist activity and neprilysin (NEP) enzymeinhibition activity. Accordingly, compounds of the invention areexpected to be useful and advantageous as therapeutic agents fortreating conditions such as hypertension and heart failure.

One aspect of the invention relates to a compound of formula I:

wherein:

r is 0, 1 or 2;

Ar is an aryl group selected from:

R¹ is selected from —COOR^(1a), —NHSO₂R^(1b), —SO₂NHR^(1d), —SO₂OH,—C(O)NH—SO₂R^(1c), —P(O)(OH)₂, —CN, —OCH(R^(1e))—COOH, tetrazol-5-yl,

R^(1a) is H, —C₁₋₆alkyl, —C₁₋₃alkylenearyl, —C₁₋₃alkyleneheteroaryl,—C₃₋₇cycloalkyl, —CH(C₁₋₄alkyl)OC(O)R^(1aa), —C₀₋₆alkylenemorpholine,

R^(1aa) is —O—C₁₋₆alkyl, —O—C₃₋₇cycloalkyl, —NR^(1ab)R^(1ac), or—CH(NH₂)CH₂COOCH₃; R^(1ab) and R^(1ac) are independently H, —C₁₋₆alkyl,or benzyl, or are taken together as —(CH₂)₃₋₆—; R^(1b) is R^(1c) or—NHC(O)R^(1c); R^(1c) is —C₁₋₆alkyl, —C₀₋₆alkylene-O—R^(lca),—C₁₋₅alkylene-NR^(1cb)R^(1cc), —C₀₋₄alkylenearyl, or—C₀₋₄alkyleneheteroaryl; R^(1ca) is H, —C₁₋₆alkyl, or—C₁₋₆alkylene-O—C₁₋₆alkyl; R^(1cb) and R^(1cc) are independently H or—C₁₋₆alkyl, or are taken together as —(CH₂)₂—O—(CH₂)₂— or—(CH₂)₂—N[C(O)CH₃]—(CH₂)₂—; R^(1d) is H, R^(1c), —C(O)R^(1c), or—C(O)NHR^(1c); R^(1e) is —C₁₋₄alkyl or aryl;

n is 0, 1, 2 or 3;

each R² is independently selected from halo, —NO₂, —C₁₋₆alkyl,—C₂₋₆alkenyl, —C₃₋₇cycloalkyl, —CN, —C(O)R^(2a), —C₀₋₅alkylene-OR^(2b),—C₀₋₅alkylene-NR^(2c)R^(2d), —C₀₋₃alkylenearyl, and—C₀₋₃alkyleneheteroaryl; R^(2a) is H, —C₁₋₆alkyl, —C₃₋₇cycloalkyl,—OR^(2b), or —NR^(2c)R^(2d); R^(2b) is H, —C₁₋₆alkyl, —C₃₋₇cycloalkyl,or —C₀₋₁alkylenearyl; and R^(2c) and R^(2d) are independently H,—C₁₋₄alkyl, or —C₀₋₁alkylenearyl;

R^(2′) is selected from H and R²;

R³ is selected from —C₁₋₁₀alkyl, —C₂₋₁₀alkenyl, —C₃₋₁₀alkynyl,—C₀₋₃alkylene-C₃₋₇cycloalkyl, —C₂₋₃alkenylene-C₃₋₇cycloalkyl,—C₂₋₃alkynylene-C₃₋₇cycloalkyl,—C₀₋₅alkylene-NR^(3a)—C₀₋₅alkylene-R^(3b),—C₀₋₅alkylene-O—C₀₋₅alkylene-R^(3b),—C₁₋₅alkylene-S—C₁₋₅alkylene-R^(3b), and —C₀₋₃alkylenearyl; R^(3a) is H,—C₁₋₆alkyl, —C₃₋₇cycloalkyl, or —C₀₋₃alkylenearyl; and R^(3b) is H,—C₁₋₆alkyl, —C₃₋₇cycloalkyl, —C₂₋₄alkenyl, —C₂₋₄alkynyl, or aryl;

X is —C₁₋₁₂alkylene-, where at least one —CH₂— moiety in the alkylene isreplaced with a —NR^(4a)—C(O)— or —C(O)—NR^(4a)— moiety, where R^(4a) isH, —OH, or —C₁₋₄alkyl;

R⁵ is selected from —C₀₋₃alkylene-SR^(5a),—C₀₋₃alkylene-C(O)NR^(5b)R^(5c), —C₀₋₃alkylene-NR^(5b)—C(O)R^(5d),—NH—C₀₋₁alkylene-P(O)(OR^(5e))₂, —C₀₋₃alkylene-P(O)OR^(5e)R^(5f),—C₀₋₂alkylene-CHR^(5g)COOH, —C₀₋₃alkylene-C(O)NR^(5h)—CHR^(5i)—COOH, and—C₀₋₃alkylene-S—SR^(5j); R^(5a) is H or —C(O)—R^(5aa); R^(5aa) is—C₁₋₆alkyl, —CO₀₋₆alkylene-C₃₋₇cycloalkyl, —C₀₋₆alkylenearyl,—C₀₋₆alkyleneheteroaryl, —C₀₋₆alkylenemorpholine,—C₀₋₆alkylenepiperazine-CH₃, —CH[N(R^(5ab))₂]-aa where aa is an aminoacid side chain, -2-pyrrolidine, —C₀₋₆alkylene-OR^(5ab),—OC₀₋₆alkylenearyl, —C₁₋₂alkylene-OC(O)—C₁₋₆alkyl,—C₁₋₂alkylene-OC(O)—C₀₋₆alkylenearyl, or —C₁₋₂alkylene-OC(O)—OC₁₋₆alkyl;R^(5ab) is independently H or —C₁₋₆alkyl; R^(5b) is H, —OH,—OC(O)R^(5ba), —CH₂COOH, —O-benzyl, -pyridyl, or —OC(S)NR^(5bb)R^(5bc);R^(5ba) is H, —C₁₋₆alkyl, aryl, —OCH₂-aryl, —CH₂O-aryl, or—NR^(5bb)R^(5bc); R^(5bb) and R^(5bc) are independently H or —C₁₋₄alkyl;R^(5c) is H, —C₁₋₆alkyl, or —C(O)—R^(5ca); R^(5ca) is H, —C₁₋₆alkyl,—C₃₋₇cycloalkyl, aryl, or heteroaryl; R^(5d) is H, —C₁₋₄alkyl,—C₀₋₃alkylenearyl, —NR^(5da)R^(5db), —CH₂SH, or —O—C₁₋₆alkyl; R^(5da)and R_(5db) are independently H or —C₁₋₄alkyl; R^(5e) is H, —C₁₋₆alkyl,—C₁₋₃alkylenearyl, —C₁₋₃alkyleneheteroaryl, —C₃₋₇cycloalkyl,—CH(CH₃)OC(O)R^(5ea),

R^(5ea) is —O—C₁₋₆alkyl, —O—C₃₋₇cycloalkyl, —NR^(5eb)R^(5ec), or—CH(NH₂)CH₂COOCH₃; R^(5eb) and R^(5ec) are independently H, —C₁₋₄alkyl,or —C₁₋₃alkylenearyl, or are taken together as —(CH₂)₃₋₆—; R^(5f) is H,—C₁₋₄alkyl, —C₀₋₃alkylenearyl, —C₁₋₃alkylene-NR^(5fa)R^(5fb), or—C₁₋₃alkylene(aryl)-C₀₋₃alkylene-NR^(5fa)R^(5fb); R^(5fa) and R^(5fb)are independently H or —C₁₋₄alkyl; R^(5g), is H, —C₁₋₆alkyl,—C₁₋₃alkylenearyl, or —CH₂—O—(CH₂)₂—OCH₃; R^(5h) is H or —C₁₋₄alkyl; andR^(5i) is H, —C₁₋₁₄alkyl, or —C₀₋₃alkylenearyl; and R^(5j) is—C₁₋₆alkyl, aryl, or —CH₂CH(NH₂)COOH;

R⁶ is selected from —C₁₋₆alkyl, —CH₂O(CH₂)₂OCH₃,—C₁₋₆alkylene-O—C₁₋₆alkyl, —C₀₋₃alkylenearyl, —C₀₋₃alkyleneheteroaryl,and —C₀₋₃alkylene-C₃₋₇cycloalkyl; and

R⁷ is H or is taken together with R⁶ to form —C₃₋₈cycloalkyl;

wherein each —CH₂— group in —(CH₂)_(r)— is optionally substituted with 1or 2 substituents independently selected from —C₁₋₄alkyl and fluoro;

each carbon atom in the alkylene moiety in X is optionally substitutedwith one or more R^(4b) groups and one —CH₂— moiety in X may be replacedwith a group selected from —C₄₋₈cycloalkylene-, —CR^(4d)═CH—, and—CH═CR^(4d)—; R^(4b) is —C₀₋₅alkylene-COOR^(4c), —C₁₋₆alkyl,—C₀₋₁alkylene-CONH₂, —C₁₋₂alkylene-OH, —C₀₋₃alkylene-C₃₋₇cycloalkyl,1H-indol-3-yl, benzyl, or hydroxybenzyl; R^(4c) is H or —C₁₋₄alkyl; andR^(4d) is —CH₂-thiophene or phenyl;

each alkyl and each aryl in R¹, R², R^(2′), R³, R^(4a-4d), and R⁵⁻⁶ isoptionally substituted with 1 to 7 fluoro atoms;

each ring in Ar and each aryl and heteroaryl in R¹, R², R^(2′), R³, andR⁵⁻⁶ is optionally substituted with 1 to 3 substituents independentlyselected from —OH, —C₁₋₆alkyl, —C₂₋₄alkenyl, —C₂₋₄alkynyl, —CN, halo,—O—C₁₋₆alkyl, —S—C₁₋₆alkyl, —S(O)—C₁₋₆alkyl, —S(O)₂—C₁₋₄alkyl, -phenyl,—NO₂, —NH₂, —NH—C₁₋₆alkyl, and —N(C₁₋₆alkyl)₂, wherein each alkyl,alkenyl and alkynyl is optionally substituted with 1 to 5 fluoro atoms;

or a pharmaceutically acceptable salt thereof.

Another aspect of the invention relates to pharmaceutical compositionscomprising a pharmaceutically acceptable carrier and a compound of theinvention. Such compositions may optionally contain other therapeuticagents such as diuretics, β₁ adrenergic receptor blockers, calciumchannel blockers, angiotensin-converting enzyme inhibitors, AT₁ receptorantagonists, neprilysin inhibitors, non-steroidal anti-inflammatoryagents, prostaglandins, anti-lipid agents, anti-diabetic agents,anti-thrombotic agents, renin inhibitors, endothelin receptorantagonists, endothelin converting enzyme inhibitors, aldosteroneantagonists, angiotensin-converting enzyme/neprilysin inhibitors,vasopressin receptor antagonists, and combinations thereof. Accordingly,in yet another aspect of the invention, a pharmaceutical compositioncomprises a compound of the invention, a second therapeutic agent, and apharmaceutically acceptable carrier. Another aspect of the inventionrelates to a combination of active agents, comprising a compound of theinvention and a second therapeutic agent. The compound of the inventioncan be formulated together or separately from the additional agent(s).When formulated separately, a pharmaceutically acceptable carrier may beincluded with the additional agent(s). Thus, yet another aspect of theinvention relates to a combination of pharmaceutical compositions, thecombination comprising: a first pharmaceutical composition comprising acompound of the invention and a first pharmaceutically acceptablecarrier; and a second pharmaceutical composition comprising a secondtherapeutic agent and a second pharmaceutically acceptable carrier. Theinvention also relates to a kit containing such pharmaceuticalcompositions, for example where the first and second pharmaceuticalcompositions are separate pharmaceutical compositions.

Compounds of the invention possess both AT₁ receptor antagonist activityand NEP enzyme inhibition activity, and are therefore expected to beuseful as therapeutic agents for treating patients suffering from adisease or disorder that is treated by antagonizing the AT₁ receptorand/or inhibiting the NEP enzyme. Thus, one aspect of the inventionrelates to a method of treating patients suffering from a disease ordisorder that is treated by antagonizing the AT₁ receptor and/orinhibiting the NEP enzyme, comprising administering to a patient atherapeutically effective amount of a compound of the invention. Anotheraspect of the invention relates to a method of treating hypertension orheart failure, comprising administering to a patient a therapeuticallyeffective amount of a compound of the invention. Still another aspect ofthe invention relates to a method for antagonizing an AT₁ receptor in amammal comprising administering to the mammal, an AT₁receptor-antagonizing amount of a compound of the invention. Yet anotheraspect of the invention relates to a method for inhibiting a NEP enzymein a mammal comprising administering to the mammal, a NEPenzyme-inhibiting amount of a compound of the invention.

Compounds of the invention that are of particular interest include thosethat exhibit an inhibitory constant (pK_(i)) for binding to an AT₁receptor greater than or equal to about 5.0; in particular those havinga pK_(i) greater than or equal to about 6.0; in one embodiment thosehaving a pK_(i) greater than or equal to about 7.0; more particularlythose having a pK_(i) greater than or equal to about 8.0; and in yetanother embodiment, those having a pK_(i) within the range of about8.0-10.0. Compounds of particular interest also include those having aNEP enzyme inhibitory concentration (pIC₅₀) greater than or equal toabout 5.0; in one embodiment those having a pIC₅₀ greater than or equalto about 6.0; in particular those having a pIC₅₀ greater than or equalto about 7.0; and most particularly those having a pIC₅₀ within therange of about 7.0-10.0. Compounds of further interest include thosehaving a pK_(i) for binding to an AT₁ receptor greater than or equal toabout 7.5 and having a NEP enzyme pIC₅₀ greater than or equal to about7.0.

Since compounds of the invention possess AT₁ receptor antagonistactivity and NEP inhibition activity, such compounds are also useful asresearch tools. Accordingly, one aspect of the invention relates to amethod of using a compound of the invention as a research tool, themethod comprising conducting a biological assay using a compound of theinvention. Compounds of the invention can also be used to evaluate newchemical compounds. Thus another aspect of the invention relates to amethod of evaluating a test compound in a biological assay, comprising:(a) conducting a biological assay with a test compound to provide afirst assay value; (b) conducting the biological assay with a compoundof the invention to provide a second assay value; wherein step (a) isconducted either before, after or concurrently with step (b); and (c)comparing the first assay value from step (a) with the second assayvalue from step (b). Exemplary biological assays include an AT₁ receptorbinding assay and a NEP enzyme inhibition assay. Still another aspect ofthe invention relates to a method of studying a biological system orsample comprising an AT₁ receptor, a NEP enzyme, or both, the methodcomprising: (a) contacting the biological system or sample with acompound of the invention; and (b) determining the effects caused by thecompound on the biological system or sample.

The invention also relates to processes and intermediates useful forpreparing compounds of the invention. In one aspect of the inventionnovel intermediates have formula III, IV, or V.

Yet another aspect of the invention relates to the use of a compound offormula I or a pharmaceutically acceptable salt thereof, for themanufacture of a medicament, especially for the manufacture of amedicament useful for treating hypertension or acute decompensated heartfailure. Another aspect of the invention relates to use of a compound ofthe invention for antagonizing an AT₁ receptor or for inhibiting a NEPenzyme in a mammal. Still another aspect of the invention relates to theuse of a compound of the invention as a research tool. Other aspects andembodiments of the invention are disclosed herein.

DETAILED DESCRIPTION OF THE INVENTION

The invention relates to compounds of formula I:

and pharmaceutically acceptable salts thereof.

As used herein, the term “compound of the invention” includes allcompounds encompassed by formula I such as the species embodied informulas Ia, Ib, Ic, II, IIa, IIb, IIc, III, IV, and V, described below.In addition, the compounds of the invention may also contain severalbasic or acidic groups (e.g., amino or carboxyl groups) and therefore,such compounds can exist as a free base, free acid, or in various saltforms. All such salt forms are included within the scope of theinvention. Finally, the compounds of the invention may also exist asprodrugs. Accordingly, those skilled in the art will recognize thatreference to a compound herein, for example, reference to a “compound ofthe invention” or a “compound of formula I” includes a compound offormula I as well as pharmaceutically acceptable salts and prodrugs ofthat compound unless otherwise indicated. Further, the term “or apharmaceutically acceptable salt and/or prodrug thereof” is intended toinclude all permutations of salts and prodrugs, such as apharmaceutically acceptable salt of a prodrug. Furthermore, solvates ofcompounds of formula I or salts thereof are included within the scope ofthe invention.

The compounds of formula I may contain one or more chiral centers andtherefore, these compounds may be prepared and used in variousstereoisomeric forms. Accordingly, the invention relates to racemicmixtures, pure stereoisomers (i.e., enantiomers or diastereomers),stereoisomer-enriched mixtures, and the like unless otherwise indicated.When a chemical structure is depicted herein without anystereochemistry, it is understood that all possible stereoisomers areencompassed by such structure. Thus, for example, the term “compound offormula I” is intended to include all possible stereoisomers of thecompound. Similarly, when a particular stereoisomer is shown or namedherein, it will be understood by those skilled in the art that minoramounts of other stereoisomers may be present in the compositions of theinvention unless otherwise indicated, provided that the utility of thecomposition as a whole is not eliminated by the presence of such otherisomers. Individual enantiomers may be obtained by numerous methods thatare well known in the art, including chiral chromatography using asuitable chiral stationary phase or support, or by chemically convertingthem into diastereomers, separating the diastereomers by conventionalmeans such as chromatography or recrystallization, then regenerating theoriginal enantiomers. Additionally, where applicable, all cis-trans orE/Z isomers (geometric isomers), tautomeric forms and topoisomeric formsof the compounds of the invention are included within the scope of theinvention unless otherwise specified.

One possible chiral center could be present in the X portion of thecompound. For example, a chiral center exists at a carbon atom in thealkylene moiety in X that is substituted with an R^(4b) group such as—C₁₋₆alkyl, for example —CH₃. This chiral center is present at thecarbon atom indicated by the symbol * in the following partial formula:

Another possible chiral center could be present in the —CR⁵R⁶R⁷ portionof the compound, when R⁶ is a group such as —C₁₋₆alkyl, for example—CH₂CH(CH₃)₂, and R⁷ is H. This chiral center is present at the carbonatom indicated by the symbol ** in the following formula:

In one embodiment of the invention, the carbon atom identified by thesymbol * and/or ** has the (R) configuration. In this embodiment,compounds of formula I have the (R) configuration at the carbon atomidentified by the symbol * and/or ** or are enriched in a stereoisomericform having the (R) configuration at this carbon atom (or atoms). Inanother embodiment, the carbon atom identified by the symbol * and/or **has the (S) configuration. In this embodiment, compounds of formula Ihave the (S) configuration at the carbon atom identified by the symbol *and/or ** or are enriched in a stereoisomeric form having the (S)configuration at this carbon atom. It is understood that a compound mayhave a chiral center at both the * and the ** carbon atoms. In suchcases, four possible diastereomers can exist. In some cases, in order tooptimize the therapeutic activity of the compounds of the invention,e.g., as hypertensive agents, it may be desirable that the carbon atomidentified by the symbol * and/or ** have a particular (R) or (S)configuration.

The compounds of the invention, as well as those compounds used in theirsynthesis, may also include isotopically-labeled compounds, i.e., whereone or more atoms have been enriched with atoms having an atomic massdifferent from the atomic mass predominately found in nature. Examplesof isotopes that may be incorporated into the compounds of formula I,for example, include, but are not limited to, ²H, ³H, ¹³C, ¹⁴C, ¹⁵N,¹⁸O, ¹⁷O, ³⁵S, ³⁶Cl, and ¹⁸F.

The compounds of formula I have been found to possess AT₁ receptorantagonizing activity and NEP enzyme inhibition activity. Among otherproperties, such compounds are expected to be useful as therapeuticagents for treating diseases such as hypertension. By combining dualactivity into a single compound, double therapy can be achieved, i.e.,AT₁ receptor antagonist activity and NEP enzyme inhibition activity canbe obtained using a single active component. Since pharmaceuticalcompositions containing one active component are typically easier toformulate than compositions containing two active components, suchsingle-component compositions provide a significant advantage overcompositions containing two active components. In addition, certaincompounds of the invention have also been found to be selective forinhibition of the AT₁ receptor over the angiotensin II type 2 (AT₂)receptor, a property that may have therapeutic advantages.

The nomenclature used herein to name the compounds of the invention isillustrated in the Examples herein. This nomenclature has been derivedusing the commercially available AutoNom software (MDL, San Leandro,Calif.).

REPRESENTATIVE EMBODIMENTS

The following substituents and values are intended to providerepresentative examples of various aspects and embodiments of theinvention. These representative values are intended to further defineand illustrate such aspects and embodiments and are not intended toexclude other embodiments or to limit the scope of the invention. Inthis regard, the representation that a particular value or substituentis preferred is not intended in any way to exclude other values orsubstituents from the invention unless specifically indicated.

In one aspect, the invention relates to compounds of formula I:

The values for r are 0, 1 or 2. In one embodiment, r is 1. Each —CH₂—group in the —(CH₂)_(r)— group may be substituted with 1 or 2substituents independently selected from —C₁₋₄alkyl (for example, —CH₃)and fluoro. In one particular embodiment, the —(CH₂)_(r)— group isunsubstituted; in another embodiment, one or two —CH₂— groups in—(CH₂)_(r)— are substituted with an —C₁₋₄alkyl group.

Ar represents an aryl group selected from:

Each ring in the Ar moiety may be substituted with 1 to 3 substituentsindependently selected from —OH, —C₁₋₆alkyl, —C₂₋₄alkenyl, —C₂₋₄alkynyl,—CN, halo, —O—C₁₋₆alkyl, —S—C₁₋₆alkyl, —S(O)—C₁₋₆alkyl,—S(O)₂—C₁₋₄alkyl, -phenyl, —NO₂, —NH₂, —NH—C₁₋₆alkyl and —N(C₁₋₆alkyl)₂.Furthermore, each of the aforementioned alkyl, alkenyl and alkynylgroups are optionally substituted with 1 to 5 fluoro atoms.

In one particular embodiment, each ring in the Ar moiety may besubstituted with 1 to 2 substituents independently selected from —OH,—C₁₋₄alkyl (for example, —CH₃), halo (for example bromo, fluoro, chloro,and di-fluoro), —O—C₁₋₄alkyl (for example, —OCH₃), and -phenyl.Exemplary substituted Ar moieties include:

Of particular interest is the embodiment where Ar is substituted with 1or 2 halo atoms.

It is understood that the Ar structure depicted as:

In one particular embodiment, Ar is an aryl group selected from:

R¹ is selected from —COOR^(1a), —NHSO₂R^(1b), —SO₂NHR^(1d), —SO₂OH,—C(O)NH—SO₂R^(1c), —P(O)(OH)₂, —CN, —OCH(R^(1e))—COOH, tetrazol-5-yl,

The R^(1a) moiety is H, —C₁₋₆alkyl, —C₁₋₃alkylenearyl,—C₁₋₃alkyleneheteroaryl, —C₃₋₇cycloalkyl, —CH(C₁₋₄alkyl)OC(O)R^(1aa),—C₀₋₆alkylenemorpholine,

R^(1aa) is —O—C₁₋₆alkyl, —O—C₃₋₇cycloalkyl, —NR^(1ab)R^(1ac), or—CH(NH₂)CH₂COOCH₃. R^(1ab) and R^(1ac) are independently selected fromH, —C₁₋₆alkyl, and benzyl, or are taken together as —(CH₂)₃₋₆—.

The R^(1b) moiety is R^(1c) or —NHC(O)R^(1c). The R^(1c) group is—C₁₋₆alkyl, —C₀₋₆alkylene-O—R^(1ca), —C₁₋₅alkylene-NR^(1cb)R^(1cc),—C₀₋₄alkylenearyl, or —C₀₋₄alkyleneheteroaryl. The R^(1ca) moiety is H,—C₁₋₆alkyl, or —C₁₋₆alkylene-O—C₁₋₆alkyl. The R^(1cb) and R^(1cc) groupsare independently selected from H and —C₁₋₆alkyl, or are taken togetheras —(CH₂)₂—O—(CH₂)₂— or —(CH₂)₂—N[C(O)CH₃]—(CH₂)₂—. The R^(1d) moiety isH, R^(1c), —C(O)R^(1c), or —C(O)NHR^(1c). The R^(1e) group is C₁₋₄alkylor aryl.

Each alkyl and each aryl in R¹ is optionally substituted with 1 to 7fluoro atoms. In addition, the term “alkyl” is intended to includedivalent alkylene groups such as those present in —C₁₋₃alkylenearyl and—C₁₋₃alkyleneheteroaryl, for example. Further, each aryl and heteroarylgroup that might be present in R¹, may be substituted with 1 to 3-OH,—C₁₋₆alkyl, —C₂₋₄alkenyl, —C₂₋₄alkynyl, —CN, halo, —O—C₁₋₆alkyl,—S—C₁₋₆alkyl, —S(O)—C₁₋₆alkyl, —S(O)₂—C₁₋₄alkyl, -phenyl, —NO₂, —NH₂,—NH—C₁₋₆alkyl, or —N(C₁₋₆alkyl)₂ groups. Further, each of theaforementioned alkyl, alkenyl and alkynyl groups may be substituted with1 to 5 fluoro atoms. It is understood that when referring to “eachalkyl,” “each aryl” and “each heteroaryl” group in R¹, the terms alsoinclude any alkyl, aryl and heteroaryl groups that might be present inthe R^(1a) through R^(1e) moieties.

In one embodiment, R¹ is —COOR^(1a) and R^(1a) is H. In anotherembodiment, R¹ is —COOR^(1a) and R^(1a) is —C₁₋₆alkyl, examples of whichinclude —CH₃, —CH₂CH₃, —(CH₂)₂CH₃, —(CH₂)₂—CF₃, —CH₂CH(CH₃)₂, —CH(CH₃)₂,—CH(CH₃)—CF₃, —CH(CH₂F)₂, —C(CH₃)₃, —(CH₂)₃CH₃, and —(CH₂)₂—CF₂CF₃.Thus, examples of R¹ include —C(O)OCH₃, —COOCH₂CH₃, —C(O)O(CH₂)₂CH₃,—C(O)OCH₂CH(CH₃)₂, —C(O)O(CH₂)₃CH₃, and so forth.

In one embodiment, R¹ is —COOR^(1a) and R^(1a) is —C₁₋₃alkylenearyl, forexample, a benzyl group, which may be substituted such as chlorobenzyl,fluorobenzyl, di fluorobenzyl, -benzyl-CH₃, -benzyl-CF₃ and-benzyl-OCF₃. Thus, examples of R¹ include: —C(O)OCH₂-benzyl,

In one embodiment, R¹ is —COOR^(1a) and R^(1a) is—C₁₋₃alkyleneheteroaryl, examples of which include —CH₂-pyridinyl. Inone embodiment, R¹ is —COOR^(1a) and R^(1a) is —C₃₋₇cycloalkyl, examplesof which include cyclopentyl.

In yet another embodiment R¹ is —COOR^(1a) and R^(1a) is—CH(C₁₋₄alkyl)OC(O)R^(1aa), where R^(1aa) is —O—C₁₋₆alkyl,—O—C₃₋₇cycloalkyl, —NR^(1ab)R^(1ac), or —CH(NH₂)CH₂COOCH₃. R^(1ab) andR^(1ac) are independently selected from H, —C₁₋₆alkyl, and benzyl, orare taken together as —(CH₂)₃₋₆—. Examples of —O—C₁₋₆alkyl groupsinclude —O—CH₂CH₃ and —O—CH(CH₃)₂. Exemplary —O—C₃₋₇cycloalkyl groupsinclude —O-cyclohexyl. Thus, examples of R¹ include—C(O)OCH(CH₃)OC(O)—O—CH₂CH₃, —C(O)OCH(CH₃)OC(O)—O—CH(CH₃)₂, and—C(O)OCH(CH₃)OC(O)—O-cyclohexyl.

In one embodiment, R¹ is —COOR^(1a) and R^(1a) is—C₀₋₆alkylenemorpholine, examples of which include —(CH₂)₂-morpholineand —(CH₂)₃-morpholine. In another embodiment, R^(1a) is

In one embodiment, R¹ is —NHSO₂R^(1b) and R^(1b) is R^(1c). The R^(1c)group is —C₁₋₆alkyl, —C₀₋₆alkylene-O—R^(1ca),—C₁₋₅alkylene-NR^(1cb)R^(1cc), —C₀₋₄alkylenearyl or—C₀₋₄alkyleneheteroaryl. The R^(1ca) moiety is H, —C₁₋₆alkyl, or—C₁₋₆alkylene-O—C₁₋₆alkyl. The R^(1cb) and R^(1cc) groups areindependently selected from H and —C₁₋₆alkyl, or are taken together as—(CH₂)₂—O—(CH₂)₂— or —(CH₂)₂—N[C(O)CH₃]—(CH₂)₂—. In one embodiment,R^(1c) is —C₁₋₆alkyl, such that exemplary R¹ groups include —NHSO₂—CH₃and the fluoro-substituted group, —NHSO₂—CF₃. In another embodiment,R^(1c) is C₀₋₄alkylenearyl, such that exemplary R¹ groups include—NHSO₂-phenyl. In another embodiment, R^(1c) is —C₀₋₄alkyleneheteroaryl,such that exemplary R¹ groups include —NHSO₂-4,5-dimethylisoxazol-3-yl.

In another embodiment, R¹ is —NHSO₂R^(1b) and R^(1b) is —NHC(O)R^(1c),where R^(1c) is defined above. In a particular embodiment, R¹ is—NHSO₂R^(1b), R^(1b) is —NHC(O)R^(1c), and R^(1c) is —C₁₋₆alkyl or—C₀₋₄alkylenearyl.

In one embodiment, R¹ is —SO₂NHR^(1d) and R^(1d) is H. In anotherembodiment, R¹ is —SO₂NHR^(1d) and R^(1d) is R^(1c), where R^(1c) isdefined above. In a particular embodiment, R^(1c) is —C₁₋₆alkyl or—C₀₋₄alkylenearyl. When R^(1c) is —C₁₋₆alkyl, exemplary R¹ groupsinclude the fluoro-substituted groups —SO₂NH—CF₃, —SO₂NH—CHF₂,—SO₂NH—CF₂CH₂F and —SO₂NH—CF₂CF₂CF₃.

In another embodiment, R¹ is —SO₂NHR^(1d) and R^(1d) is —C(O)R^(1c),where R^(1c) is defined above. In a particular embodiment, R^(1c) is—C₁₋₆alkyl or —C₀₋₄alkylenearyl. When R^(1c) is —C₁₋₆alkyl, exemplary R¹groups include —SO₂NHC(O)CH₃ and —SO₂NHC(O)(CH₂)₂CH₃. When R^(1c) is—C₀₋₆alkylene-O—R^(1ca) and R^(1ca) is H, exemplary R¹ groups include—SO₂NHC(O)CH₂OH, —SO₂NHC(O)CH(CH₃)OH, and —SO₂NHC(O)C(CH₃)₂OH. WhenR^(1c) is —C₀₋₆alkylene-O—R^(1ca) and R^(1ca) is —C₁₋₆alkyl, exemplaryR¹ groups include —SO₂NHC(O)CH₂—O—CH₃, —SO₂NHC(O)—O—CH₃, and—SO₂NHC(O)—O—CH₂CH₃. When R^(1c) is —C₀₋₆alkylene-O—R^(1ca) and R^(1ca)is —C₁₋₆alkylene-O—C₁₋₆alkyl, exemplary R¹ groups include—SO₂NHC(O)CH₂—O—(CH₂)₂—O—CH₃. When R^(1c) is—C₁₋₅alkylene-NR^(1cb)R^(1cc), exemplary R¹ groups include—SO₂NHC(O)CH₂N(CH₃)₂, —SO₂NHC(O)CH₂NH₂, and —SO₂NHC(O)—CH(CH₃)NH₂.Another example when R^(1c) is —C₁₋₅alkylene-NR^(1cb)R^(1cc) is wherethe R^(1cb) and R^(1cc) are taken together as —(CH₂)₂—O—(CH₂)₂— or—(CH₂)₂—N[C(O)CH₃]—(CH₂)₂—. Such exemplary R¹ groups include:

In another embodiment, R¹ is —SO₂NHR^(1d) and R^(1d) is —C(O)NHR^(1c),where R^(1c) is defined above. In a particular embodiment, R^(1c) is—C₁₋₆alkyl or —C₀₋₄alkylenearyl. When R^(1c) is —C₁₋₆alkyl, exemplary R¹groups include —SO₂NHC(O)NH—CH₂CH₃ and —SO₂NHC(O)NH—(CH₂)₂CH₃. WhenR^(1c) is —C₀₋₄alkylenearyl, exemplary R¹ groups include—SO₂NHC(O)NH-phenyl.

In yet another embodiment, R¹ is —SO₂OH. In one embodiment, —P(O)(OH)₂.In still another embodiment, R¹ is —CN.

In another embodiment, R¹ is —C(O)NH—SO₂R^(1c), where R^(1c) is definedabove. In a particular embodiment, R^(1c) is —C₁₋₆alkyl or—C₀₋₄alkylenearyl. When R^(1c) is —C₁₋₆alkyl, exemplary R¹ groupsinclude —C(O)—NH—SO₂—CH₃, —C(O)—NH—SO₂—CH₂CH₃ and the fluoro-substituted—C(O)—NH—SO₂—CF₃ group.

In another embodiment, R¹ is —OCH(R^(1e))—COOH, where R^(1e) is—C₁₋₄alkyl or aryl. Examples of such R¹ groups include, —OCH(CH₃)—COOHand —OCH(phenyl)-COOH.

In one particular embodiment, R¹ is selected from —COOR^(1a) andtetrazol-5-yl. In another embodiment, R¹ is —COOR^(1a) and R^(1a) is Hor —C₁₋₆alkyl.

The values for n are 0, 1, 2 or 3. In one embodiment, n is 0. In anotherembodiment, n is 1.

Each R² is independently selected from halo, —NO₂, —C₁₋₆alkyl,—C₂₋₆alkenyl, —C₃₋₇cycloalkyl, —CN, —C(O)R^(2a), —C₀₋₅alkylene-OR^(2b),—C₀₋₅alkylene-NR^(2c)R^(2d), —C₀₋₃alkylenearyl, and—C₀₋₃alkyleneheteroaryl. The R^(2a) moiety is H, —C₁₋₆alkyl,—C₃₋₇cycloalkyl, —OR^(2b), or —NR^(2c)R^(2d). R^(2b) is selected from H,—C₁₋₆alkyl, —C₃₋₇cycloalkyl, and —C₀₋₁alkylenearyl; and R^(2c) andR^(2d) are independently selected from H, —C₁₋₄alkyl, and—C₀₋₁alkylenearyl.

In one particular embodiment, R² is halo, for example, chloro. In yetanother embodiment, R² is —C₁₋₆alkyl such as —CH₃, andfluoro-substituted alkyl groups such as —CH₂F and —CF₃. In anotherembodiment R² is —C₀₋₅alkylene-OR^(2b) and R^(2b) is H; one such R²group is —CH₂OH.

The R^(2′) moiety can be H or a moiety as defined above for R², i.e., isselected from, halo, —NO₂, —C₁₋₆alkyl, —C₂₋₆alkenyl, —C₃₋₇cycloalkyl,—CN, —C(O)R^(2a), —C₀₋₅alkylene-OR^(2b), —C₀₋₅alkylene-NR^(2c)R^(2d),—C₀₋₃alkylenearyl, and —C₀₋₃alkyleneheteroaryl. In one particularembodiment, R^(2′) is H. In another embodiment R^(2′) is —C(O)R^(2a) andR^(2a) is —C₁₋₆alkyl such as —CH₃.

Each alkyl and each aryl in R² and R^(2′) is optionally substituted with1 to 7 fluoro atoms. It is understood that when referring to the “alkyl”in R² or R^(2′), the term includes any alkyl groups that might bepresent in the R^(2a), R^(2b), R^(2c) and R^(2d) moieties. In addition,each aryl and heteroaryl in R² and R^(2′), for example in—C₀₋₃alkylenearyl and —C₀₋₃alkyleneheteroaryl, may be substituted with 1to 3-OH, —C₁₋₆alkyl, —C₂₋₄alkenyl, —C₂₋₄alkynyl, —CN, halo,—O—C₁₋₆alkyl, —S—C₁₋₆alkyl, —S(O)—C₁₋₆alkyl, —S(O)₂—C₁₋₄alkyl, -phenyl,—NO₂, —NH₂, —NH—C₁₋₆alkyl, or —N(C₁₋₆alkyl)₂ groups. Further, each ofthe aforementioned alkyl, alkenyl and alkynyl groups may be substitutedwith 1 to 5 fluoro atoms. It is understood that when referring to the“aryl” or the “heteroaryl” in R² or R^(2′), the terms includes any arylor heteroaryl groups that might be present in the R^(2a), R^(2b), R^(2c)and R^(2d) moieties.

R³ is selected from —C₁₋₁₀alkyl, —C₂₋₁₀alkenyl, —C₃₋₁₀alkynyl,—C₀₋₃alkylene-C₃₋₇cycloalkyl, —C₂₋₃alkenylene-C₃₋₇cycloalkyl,—C₂₋₃alkynylene-C₃₋₇cycloalkyl,—C₀₋₅alkylene-NR^(3a)C₀₋₅alkylene-R^(3b),—C₀₋₅alkylene-O—C₀₋₅alkylene-R^(3b),—C₁₋₅alkylene-S—C₁₋₅alkylene-R^(3b), and —C₀₋₃alkylenearyl (for example,—C₀₋₁alkylenearyl such as phenyl and benzyl). The R^(3a) moiety is H,—C₁₋₆alkyl, —C₃₋₇cycloalkyl, or —C₀₋₃alkylenearyl (for example,—C₀₋₁alkylenearyl such as phenyl and benzyl). R^(3b) is H, —C₁₋₆alkyl,—C₃₋₇cycloalkyl, —C₂₋₄alkenyl, —C₂₋₄alkynyl, or aryl (such as phenyl).

In addition, each alkyl and each aryl in R³ is optionally substitutedwith 1 to 7 fluoro atoms, where the term “alkyl” is intended to includedivalent alkylene groups such as those present in—C₀₋₃alkylene-C₃₋₇cycloalkyl and —C₀₋₃alkylenearyl, for example. Eacharyl in R³, for example in —C₀₋₃alkylenearyl or aryl, may be substitutedwith 1 to 3-OH, —C₁₋₆alkyl, —C₂₋₄alkenyl, —C₂₋₄alkynyl, —CN, halo,—O—C₁₋₆alkyl, —S—C₁₋₆alkyl —S(O)—C₁₋₆alkyl, —S(O)₂—C₁₋₄alkyl, -phenyl,—NO₂, —NH₂, —NH—C₁₋₆alkyl, or —N(C₁₋₆alkyl)₂ groups. Further, each ofthe aforementioned alkyl, alkenyl and alkynyl groups may be substitutedwith 1 to 5 fluoro atoms. It is understood that when referring to “eachalkyl” and “each aryl” group in R³, the terms also include any alkyl andaryl groups that might be present in the R^(3a) and R^(3b) moieties.

In one embodiment, R³ is —C₁₋₁₀alkyl optionally substituted with 1 to 7fluoro atoms. In another embodiment, R³ is —C₂₋₇alkyl; and in yetanother embodiment, R³ is —C₃₋₅alkyl. Examples of such R³ groupsinclude, —CH₃, —CF₃, —CH₂CH₃, —(CH₂)₂CH₃, —(CH₂)₃CH₃, —CH₂—CH(CH₃)₂,—CH₂—CH(CH₃)CH₂CH₃, —(CH₂)₂—CH(CH₃)₂, —CH(CH₂CH₃)₂, and —(CH₂)₄CH₃.

In another embodiment, R³ is —C₂₋₁₀alkenyl such as —CH₂CH═CHCH₃. In yetanother embodiment, R³ is —C₃₋₁₀alkynyl such as —CH₂C≡CCH₃.

In another embodiment, R³ is —C₀₋₃alkylene-C₃₋₇cycloalkyl such as-cyclopropyl, —CH₂-cyclopropyl, cyclopentyl, —CH₂-cyclopentyl,—(CH₂)₂-cyclopentyl, and —CH₂-cyclohexyl. In a particular embodiment, R³is —C₀₋₁alkylene-C₃₋₅cycloalkyl. In one embodiment, R³ is—C₂₋₃alkenylene-C₃₋₇cycloalkyl, such as —CH₂CH═CH-cyclopentyl; and inanother embodiment, R³ is —C₂₋₃alkynylene-C₃₋₇cycloalkyl, such as—CH₂C≡C-cyclopentyl.

In yet another embodiment, R³ is—C₀₋₅alkylene-NR^(3a)—C₀₋₅alkylene-R^(3b). In one particular embodiment,R^(3a) is H and R^(3b) is —C₁₋₆alkyl. Examples of such R³ groups include—NHCH₂CH₃, —NHCH(CH₃)₂, —NH(CH₂)₂CH₃, —NH(CH₂)₃CH₃, —NHCH(CH₃)CH₂CH₃,—NH(CH₂)₄CH₃, and —NH(CH₂)₅CH₃.

In another embodiment, R³ is —C₀₋₅alkylene-O—C₁₋₅alkylene. In oneparticular embodiment R^(3b) is selected from H, —C₁₋₆alkyl, and aryl.Examples of such R³ groups include —OCH₃, —O—CH₂CH₃, —OCH(CH₃)₂,—O(CH₂)₂CH₃, —O(CH₂)₃CH₃, —OCH₂CH(CH₃)₂, —O-phenyl, and —O-benzyl. Inanother embodiment, R³ is —C₀₋₅alkylene-O—C₀₋₅alkylene-R^(3b), whereR^(3b) is —C₁₋₆alkyl, and in another embodiment, R³ is —O—C₁₋₅alkyl.

In another embodiment, R³ is —C₁₋₅alkylene-S—C₁₋₅alkylene-R^(3b), and inone particular embodiment R^(3b) is H, such as when R³ is —CH₂—S—CH₂CH₃.In another embodiment, R³ is —C₀₋₃alkylenearyl, such as phenyl, benzyl,and —(CH₂)₂-phenyl.

X is —C₁₋₁₂alkylene-, where at least one —CH₂— moiety in the alkylene isreplaced with a —NR^(4a)—C(O)— or —C(O)—NR^(4a)— moiety. Thus X can be—C₁alkylene-, —C₁alkylene-, —C₂alkylene-, —C₃alkylene-, —C₄alkylene-,—C₅alkylene-, —C₆alkylene-, —C₇alkylene-, —C₈alkylene, —C₉alkylene-,—C₁₀alkylene-, —C₁₁alkylene-, or —C₁₂alkylene-, with at least one —CH₂—moiety being replaced. R^(4a) is selected from H, —OH, and —C₁₋₄alkyl.In one embodiment, R^(4a) is H. Each carbon atom in the—C₁₋₁₂alkylene-moiety may be substituted with one or more R^(4b) groups.R^(4b) is selected from —C₀₋₅alkylene-COOR^(4c), —C₁₋₆alkyl,—C₀₋₁alkylene-CONH₂, —C₁₋₂alkylene-OH, —C₀₋₃alkylene-C₃₋₇cycloalkyl,1H-indol-3-yl, benzyl, and hydroxybenzyl, where R^(4c) is H or—C₁₋₄alkyl.

In one embodiment, the carbon atoms in —C₁₋₂alkylene- are unsubstituted,i.e., there are no R^(4b) groups. In another embodiment, one carbon atomis substituted with one R^(4b) group; and in another embodiment, 1 or 2carbon atoms are substituted with one or two R^(4b) groups. In oneembodiment, R^(4b) is —C₀₋₅alkylene-COOR^(4c), where R^(4c) is H or—C₁₋₄alkyl. Examples of such R^(4b) groups include —COOH, —CH₂COOH,—(CH₂)₂COOH, and CH₂COOCH₃. In another embodiment, R^(4b) is —C₁₋₆alkyl,for example —CH₃ or —CH(CH₃)₂. In one embodiment, R^(4b) is—C₀₋₁alkylene-CONH₂, for example —CH₂—CONH₂ or —(CH₂)₂—CONH₂. In yetanother embodiment, R^(4b) is —C₁₋₂alkylene-OH, for example CH₂—OH. Inone embodiment, R^(4b) is 1H-indol-3-yl, benzyl, or hydroxybenzyl.

In addition, one —CH₂— moiety in X may be replaced with a group selectedfrom —C₄₋₈cycloalkylene-, —CR^(4d)═CH—, and —CH═CR^(4d)—. R^(4d) isselected from —CH₂-thiophene and phenyl. In one embodiment, none of the—CH₂— moieties are so replaced. In another embodiment, one —CH₂— moietyin X is replaced with —C₄₋₈cycloalkylene-, for example, cyclohexylene.In another embodiment, one —CH₂— moiety is replaced with —CH═CR^(4d)—,where R^(4d) is —CH₂-thiophene such as —CH₂-thiophen-2-yl.

Each alkyl and each aryl in R^(4a), R^(4b), R^(4c), and R^(4d), may besubstituted with 1 to 7 fluoro atoms, and the term “alkyl” is intendedto include divalent alkylene groups such as that present in—C₀₋₅alkylene-COOR^(4c), for example. It is noted that the R^(4b) group,—C₀₋₃alkylene-C₃₋₇cycloalkyl moiety is intended to include a—C₃₋₇cycloalkyl linked to the X —C₁₋₁₂alkylene-chain by a bond as wellas a —C₃₋₇cycloalkyl that is directly attached to the chain, asillustrated below:

In one embodiment, one to four —CH₂— moieties are replaced with—NR^(4a)—C(O)— or —C(O)—NR^(4a)— moieties; and in another embodiment one—CH₂— moiety is replaced, examples of which include: —C(O)NH—, —NHC(O)—,and —CH₂—NHC(O)—. In one embodiment, X is —C₁₋₆alkylene- and one to four—CH₂— moieties are replaced with a —NR^(4a)—C(O)— or —C(O)—NR^(4a)—moiety; and in another embodiment X is —C₁₋₄alkylene- and one or two—CH₂-moieties are replaced. In one embodiment X is —C₁₋₂alkylene- andone —CH₂— moiety is replaced. In another embodiment X is —C₁alkylene-and one —CH₂— moiety is replaced, i.e., X is —NR^(4a)—C(O)— or—C(O)—NR^(4a)—, for example, —C(O)—NH—. When more than one —CH₂— moietyin —C₁₋₁₂alkylene- is replaced with a —NR^(4a)—C(O)— or —C(O)—NR^(4a)—moiety, the replaced moieties may be contiguous or non-contiguous. Inone particular embodiment, the replaced moieties are contiguous.Exemplary X groups include the following, which depict: examples whereone or more —CH₂— moieties are replaced with —NR^(4a)—C(O)— or—C(O)—NR^(4a)— moieties; examples where —CH₂— moieties are replaced witha group selected from —C₄₋₈cycloalkylene-, —CR^(4d)═CH—, and—CH═CR^(4d)—; and examples where carbon atoms in the —C₁₋₁₂alkylene-group are substituted with one or more R^(4b) groups:

—C₁alkylene- with one —CH₂— moiety replaced:

-   -   —C(O)NH—    -   —NHC(O)—

—C₂alkylene- with one —CH₂— moiety replaced:

-   -   —CH₂—NHC(O)—    -   —C(O)NH—CH₂—    -   —CH₂—C(O)NH—    -   —CH[CH(CH₃)₂]—C(O)NH—    -   —CH(COOH)—NHC(O)—    -   —CH(CH₂COOH)—NHC(O)—    -   —CH[(CH₂)₂COOH]—NHC(O)—    -   —CH(CH₂COOCH₃)—NHC(O)—    -   —CH(CH₃)—NHC(O)—    -   —CH(CH(CH₃)₂)—NHC(O)—    -   —CH(CH₂—CONH₂)—NHC(O)—    -   —CH[(CH₂)₂—CONH₂]—NHC(O)—    -   —CH(CH₂—OH)—NHC(O)—    -   —CH(benzyl)-NHC(O)—    -   —CH(4-hydroxybenzyl)-NHC(O)—    -   —CH(1H-indol-3-yl)-NHC(O)—

—C₂alkylene- with two —CH₂— moieties replaced:

-   -   —C(O)NH—NHC(O)—    -   —CH═C(—CH₂-thiopheny-2-yl)-C(O)NH—

—C₃alkylene- with one —CH₂— moiety replaced:

-   -   —(CH₂)₂—NHC(O)—    -   —(CH₂)₂—C(O)NH—    -   —CH(CH₃)—CH₂—NHC(O)—    -   —CH[CH(CH₃)₂]—CH₂—NHC(O)—    -   —CH(COOH)—CH₂—NHC(O)—    -   —CH₂—CH(COOH)—NHC(O)—    -   —CH₂—C(CH₃)₂—NHC(O)—

—C₃alkylene- with two —CH₂— moieties replaced:

-   -   —NHC(O)—CH₂—NHC(O)—

—C₄alkylene- with one —CH₂— moiety replaced:

-   -   —(CH₂)₃—NHC(O)—    -   —C(O)NH—CH₂—CH(COOH)—CH₂

—C₄alkylene- with two —CH₂— moieties replaced:

-   -   —C(O)NH—CH(benzyl)-CH₂—NHC(O)—    -   —C(O)NH—CH(benzyl)-CH₂—C(O)NH—    -   —CH₂—NHC(O)—CH₂—NHC(O)—    -   —CH(benzyl)-NHC(O)—CH₂—NHC(O)—    -   —CH(1H-indol-3-yl)-NHC(O)—CH₂—NHC(O)—

—C₄alkylene- with three —CH₂— moieties replaced:

-   -   —CH₂—NHC(O)-cyclohexylene-NHC(O)—    -   —CH₂—N(OH)C(O)-cyclohexylene-NHC(O)—

—C₅alkylene- with two —CH₂— moieties replaced:

-   -   —CH₂—NHC(O)—CH₂—CH(COOH)—NHC(O)—    -   —CH₂—NHC(O)—(CH₂)₂—NHC(O)—    -   —C(O)NH—(CH₂)₂—C(O)N(OH)—CH₂    -   —C(O)NH—(CH₂)₂—CH(COOH)—NHC(O)—    -   —CH(COOH)—CH₂—NHC(O)—CH₂—NHC(O)—    -   —(CH₂)₂—NHC(O)-cyclohexylene-NHC(O)—    -   —CH₂—CH(COOH)—NHC(O)—CH₂—NHC(O)—

—C₆alkylene- with two —CH₂— moieties replaced:

-   -   —C(O)NH—(CH₂)₄—NHC(O)—    -   —CH₂—NHC(O)—(CH₂)₂—CH(COOH)—NHC(O)—    -   —C(O)NH—(CH₂)₃—CH(COOH)—NHC(O)—

—C₆alkylene- with three —CH₂— moieties replaced:

-   -   —C(O)NH—(CH₂)₂—NHC(O)—CH₂—NHC(O)—

—C₆alkylene- with four —CH₂— moieties replaced:

-   -   —C(O)NH—(CH₂)₂—NHC(O)-cyclohexylene-NHC(O)—

—C₇alkylene- with two —CH₂— moieties replaced:

-   -   —CH₂—NHC(O)—(CH₂)₄—NHC(O)—    -   —C(O)NH—(CH₂)₄—CH(COOH)—NHC(O)—

—C₇alkylene- with three —CH₂— moieties replaced:

-   -   —CH[CH(CH₃)₂]—C(O)NH—(CH₂)₂—NHC(O)—CH₂—NHC(O)—

—C₇alkylene- with four —CH₂— moieties replaced:

-   -   —CH₂—NHC(O)—(CH₂)₂—NHC(O)-cyclohexylene-NHC(O)—    -   —CH₂—C(O)NH—(CH₂)₂—NHC(O)-cyclohexylene-NHC(O)—

—C₈alkylene- with three —CH₂— moieties replaced:

-   -   —C(O)NH—(CH₂)₄—NHC(O)—CH₂—NHC(O)—

—C₈alkylene- with four —CH₂— moieties replaced:

-   -   —C(O)NH—(CH₂)₄—NHC(O)-cyclohexylene-NHC(O)—

—C₉alkylene- with two —CH₂— moieties replaced:

-   -   —CH₂—NHC(O)—(CH₂)₆—NHC(O)—

—C₉alkylene- with four —CH₂— moieties replaced:

-   -   —CH₂—NHC(O)—(CH₂)₄—NHC(O)-cyclohexylene-NHC(O)—

—C₁₀alkylene- with four —CH₂— moieties replaced:

-   -   —C(O)NH—(CH₂)₆—NHC(O)-cyclohexylene-NHC(O)—

—C₁₁alkylene- with three —CH₂— moieties replaced:

-   -   —CH(CH(CH₃)₂)—C(O)NH—(CH₂)₆—NHC(O)—CH₂—NHC(O)—

—C₁₁alkylene- with four —CH₂— moieties replaced:

-   -   —CH₂—NHC(O)—(CH₂)₆—NHC(O)-cyclohexylene-NHC(O)—        In one particular embodiment, X is —C₁₋₆alkylene- with one or        two —CH₂— moieties being replaced with —NHC(O)— or —C(O)NH—, and        in another embodiment X is —C₁₋₄alkylene- with one or two —CH₂—        moieties being replaced. In another embodiment, X is selected        from —C(O)NH—, —NHC(O)—, and —CH₂—NHC(O)—. In yet another        embodiment, X is —C(O)NH—.

R⁵ is selected from —C₀₋₃alkylene-SR^(5a),—C₀₋₃alkylene-C(O)NR^(5b)R^(5c), —C₀₋₃alkylene-NR^(5b)—C(O)R^(5d),—NH—C₀₋₁alkylene-P(O)(OR^(5e))₂, —C₀₋₃alkylene-P(O)OR^(5e)R^(5f),—C₀₋₂alkylene-CHR^(5g)COOH, —C₀₋₃alkylene-C(O)NR^(5h)—CHR^(5i)—COOH, and—C₀₋₃alkylene-S—SR^(5j). Each alkyl and each aryl in R⁵ is optionallysubstituted with 1 to 7 fluoro atoms, where the term “alkyl” is intendedto include divalent alkylene groups such as those present in—C₀₋₃alkylene-SR^(5a) and —C₀₋₃alkylene-P(O)OR^(5e)R^(5f), for example.Each aryl and heteroaryl in R⁵ may be substituted with 1 to 3-OH,—C₁₋₆alkyl, —C₂₋₄alkenyl, —C₂₋₄alkynyl, —CN, halo, —O—C₁₋₆alkyl,—S—C₁₋₆alkyl, —S(O)—C₁₋₆alkyl, —S(O)₂—C₁₋₄alkyl, -phenyl, —NO₂, —NH₂,—NH—C₁₋₆alkyl, or —N(C₁₋₆alkyl)₂ groups. Further, each of theaforementioned alkyl, alkenyl and alkynyl groups may be substituted with1 to 5 fluoro atoms. It is understood that when referring to “eachalkyl,” “each aryl” and “each heteroaryl” group in R⁵, the terms alsoinclude any alkyl, aryl and heteroaryl groups that might be present inthe R^(5a-5j), R^(5aa), R^(5ab), R^(5ba), R^(5bb), R^(5bc), R^(5ca),R^(5da), R^(5db), R^(5ea), R^(5eb), R^(5ec), R^(5fa), R^(5fb) moieties.

In one embodiment, R⁵ is —C₀₋₃alkylene-SR^(5a). R^(5a) is H or—C(O)—R^(5aa). The R^(5aa) group is —C₁₋₆alkyl,—C₀₋₆alkylene-C₃₋₇cycloalkyl, —C₀₋₆alkylenearyl,—C₀₋₆alkyleneheteroaryl, —C₀₋₆alkylenemorpholine,—C₀₋₆alkylenepiperazine-CH₃, —CH[N(R^(5ab))₂]-aa where aa is an aminoacid side chain, -2-pyrrolidine, —C₀₋₆alkylene-OR^(5ab),—OC₀₋₆alkylenearyl, —C₁₋₁₂alkylene-OC(O)—C₁₋₆alkyl,—C₁₋₂alkylene-OC(O)—C₀₋₆alkylenearyl, or —C₁₋₂alkylene-OC(O)—OC₁₋₆alkyl.The R^(5ab) group is independently H or —C₁₋₆alkyl. In one specificembodiment, R^(5a) is H, for example R⁵ can be —SH or —CH₂SH. In anotherembodiment, R^(5a) is —C(O)—R^(5aa), and R^(5aa)—C₁₋₆alkyl. Exemplary—C₁₋₆alkyl groups include —CH₃, —CH₂CH₃, —CH(CH₃)₂, —C(CH₃)₃, and—CH₂CH(CH₃)₂. Thus, examples of R⁵ include —SC(O)CH₃, —CH₂SC(O)CH₃,—CH₂SC(O)CH₂CH₃, —CH₂SC(O)CH(CH₃)₂, —CH₂SC(O)C(CH₃)₃, and—CH₂SC(O)CH₂CH(CH₃)₂. In one embodiment, R^(5a) is selected from H and—C(O)—C₁₋₆alkyl.

In one embodiment, R⁵ is —C₀₋₃alkylene-SR^(5a), where R^(5a) is—C(O)—R^(5aa), and R^(5aa) is —C₀₋₆alkylene-C₃₋₇cycloalkyl. ExemplaryC₃₋₇cycloalkyl groups include cyclopentyl and cyclohexyl. Thus, examplesof R⁵ include —CH₂SC(O)-cyclopentyl, —CH₂SC(O)-cyclohexyl, and—CH₂SC(O)—CH₂-cyclopentyl. In another embodiment, R⁵ is—C₀₋₃alkylene-SR^(5a), where R^(5a) is —C(O)—R^(5aa), and R^(5aa) is—C₀₋₆alkylenearyl. In one specific embodiment, the aryl is optionallysubstituted with 1 to 3 substituents such as —O—C₁₋₆alkyl. Exemplaryaryl groups include phenyl and -phenyl-OCH₃. Thus, examples of R⁵include —CH₂SC(O)-phenyl and —CH₂SC(O)-phenyl-OCH₃. In yet anotherembodiment, R⁵ is —C₀₋₃alkylene-SR^(5a), where R^(5a) is —C(O)—R^(5aa),and R^(5aa) is —C₀₋₆alkyleneheteroaryl. Exemplary heteroaryl groupsinclude furanyl, thienyl and pyridinyl. Thus, examples of R⁵ include:—CH₂SC(±)-2-pyridine, —CH₂SC(±)-3-pyridine, and —CH₂SC(±)-4-pyridine.

In another embodiment, is —C₀₋₃alkylene-SR^(5a), where R^(5a) is—C(O)—R^(5aa), and R^(5aa) is —C₀₋₆alkylenemorpholine:

more particularly, —C(O)—C₁₋₃alkylenemorpholine. Thus, examples of R⁵include —CH₂S—C(O)CH₂-morpholine and —CH₂S—C(O)(CH₂)₂-morpholine. Inanother embodiment, R⁵ is —C₀₋₃alkylene-SR^(5a), where R^(5a) is—C(O)—R^(5aa), and R^(5aa) is —C₀₋₆alkylenepiperazine-CH₃. Thus,examples of R⁵ include —CH₂S—C(O)(CH₂)₂-piperazine-CH₃.

In one embodiment, R⁵ is —C₀₋₃alkylene-SR^(5a), where R^(5a) is—C(O)—R^(5aa), and R^(5aa) is —CH[N(R^(5ab))₂]-aa where aa is an aminoacid side chain. For example, the amino acid side chain could be—CH(CH₃)₂, the valine side chain, —CH₂CH(CH₃)₂ (leucine side chain),—CH(CH₃)CH₂CH₃ (isoleucine side chain), —CH₂COOH (aspartic acid sidechain), —(CH₂)₂COOH (glutamic acid side chain), —CH(OH)(CH₃) (threonineside chain), -benzyl (phenylalanine side chain), -4-hydroxybenzyl(tyrosine side chain), and —(CH₂)₂SCH₃ (methionine side chain). Thus,examples of R⁵ include —CH₂S—C(O)—CH(NH₂)—CH(CH₃)₂,—CH₂SC(O)CH(NH₂)—CH₂CH(CH₃)₂, —CH₂SC(O)CH(NH₂)—CH(CH₃)CH₂CH₃,—CH₂SC(O)CH(NH₂)—CH₂COOH, —CH₂SC(O)CH(NH₂)—(CH₂)₂COOH,—CH₂SC(O)CH(NH₂)—CH(OH)(CH₃), —CH₂SC(O)—CH(NH₂)-benzyl,—CH₂SC(O)CH(NH₂)-4-hydroxybenzyl, and —CH₂SC(O)CH(NH₂)—(CH₂)₂SCH₃.

In yet another embodiment, R⁵ is —C₀₋₃alkylene-SR^(5a), where R^(5a) is—C(O)—R^(5aa), and R^(5aa) is −2-pyrrolidine:

Thus, an example of R⁵ is —CH₂S—C(±)-2-pyrrolidine.

In yet another embodiment, R⁵ is —C₀₋₃alkylene-SR^(5a), where R^(5a) is—C(O)—R^(5aa), and R^(5aa) is —C₀₋₆alkylene-OR^(5ab). In one embodiment,R^(5ab) is H, such that R^(5a) is —C(O)—C₀₋₆alkylene-OH. In anotherembodiment, R^(5ab) is —C₁₋₆alkyl, such that R^(5a) is—C(O)—C₀₋₆alkylene-OC₁₋₆alkyl, for example, R⁵ can be —CH₂SC(O)—OCH₂CH₃.In yet another embodiment, R⁵ is —C₀₋₃alkylene-SR^(5a), where R^(5a) is—C(O)—R^(5aa), and R^(5aa) is —OC₀₋₆alkylenearyl. In yet anotherembodiment, R⁵ is —C₀₋₃alkylene-SR^(5a), where R^(5a) is —C(O)—R^(5aa),and R^(5aa) is —C₁₋₂alkylene-OC(O)—C₁₋₆alkyl; and in another embodiment,R⁵ is —C₀₋₃alkylene-SR^(5a), where R^(5a) is —C(O)—R⁵aa, and R^(5aa) is—C₁₋₂alkylene-OC(O)—C₀₋₆alkylenearyl; and in still another embodiment,R⁵ is —C₀₋₃alkylene-SR^(5a), where R^(5a) is —C(O)—R^(5aa), and R^(5aa)is —O—C₁₋₂alkylene-OC(O)—OC₁₋₆alkyl, for example, R⁵ is—CH₂SC(O)O—CH(CH₃)OC(O)OCH(CH₃)₂.

In one embodiment, R⁵ is —C₀₋₃alkylene-C(O)NR^(5b)R^(5c). The R^(5b)moiety is H, —OH, —OC(O)R^(5ba), —CH₂COOH, —O-benzyl, -pyridyl, or—OC(S)NR^(5bb)R^(5bc). R^(5ba) is H, —C₁₋₆alkyl, aryl, —OCH₂-aryl (forexample, —OCH₂-phenyl), —CH₂O-aryl (for example, —CH₂O-phenyl), or—NR^(5bb)R^(5bc). The R^(5bb) and R^(5bc) moieties are independentlyselected from H and —C₁₋₄alkyl. In one embodiment, R^(5b) is —OH or—OC(O)R^(5ba), where —R^(5ba) is —C₁₋₆alkyl. R^(5c) is H, —C₁₋₆alkyl, or—C(O)R^(5ca), where R^(5ca) is H, —C₁₋₆alkyl, —C₃₋₇cycloalkyl, aryl, orheteroaryl. In one embodiment, R⁵ is —C₀₋₃alkylene-C(O)NR^(5b)R^(5c) andR^(5c) is H. In one specific embodiment, R⁵ is—C₀₋₃alkylene-C(O)NR^(5b)R^(5c), where R^(5b) is —OH and R^(5c) is H,for example, R⁵ can be —C(O)N(OH)H or —CH₂C(O)N(OH)H. In anotherembodiment, R⁵ is —C₀₋₃alkylene-c(O)NR^(5b)R^(5c), where R^(5b) is—OC(O)R^(5ba), R^(5ba) is —C₁₋₆alkyl, and R^(5c) is H Thus, examples ofR⁵ include —C(O)N[OC(O)CH₃]H and —C(O)N[OC(O)C(CH₃)₃]H. In still anotherembodiment, R⁵ is —C₀₋₃alkylene-C(O)NR^(5b)R^(5c) and both R^(5b) andR^(5c) are H, for example, R⁵ can be —C(O)NH₂. In yet anotherembodiment, R⁵ is —C₀₋₃alkylene-C(O)NR^(5b)R^(5c), where R^(5b) is—OC(O)R^(5ba), R^(5ba) is —OCH₂-aryl or —CH₂O-aryl, and R^(5c) is H.Thus, examples of R⁵ include —CH₂—C(O)NH[OC(O)OCH₂-phenyl] and—CH₂—C(O)N[OC(O)CH₂O-phenyl]H. In another embodiment, R⁵ is—C₀₋₃alkylene-C(O)NR^(5b)R^(5c), where R^(5b) is —OC(S)NR^(5bb)R^(5bc),R^(5bb) and R^(5bc) are both —C₁₋₄alkyl, and R^(5c) is H, for example,R⁵ can be —CH₂—C(O)N[OC(S)N(CH₃)₂]H. In another embodiment, R⁵ is—C₀₋₃alkylene-C(O)NR^(5b)R^(5c), where R^(5b) is —CH₂COOH and R^(5c) isH, for example, R⁵ can be —C(O)NH—(CH₂COOH).

In one embodiment, R⁵ is —C₀₋₃alkylene-NR^(5b)—C(O)R^(5d). The R^(5d)moiety is H, —C₁₋₄alkyl, —C₀₋₃alkylenearyl, —NR^(5da)R^(5db), —CH₂SH, or—O—C₁₋₆alkyl. The R^(5da) and R^(5db) moieties are independentlyselected from H and —C₁₋₄alkyl. In one embodiment, R^(5b) is —OH andR^(5d) is H, for example, R⁵ can be —CH₂—N(OH)C(O)H; and in anotherembodiment, R^(5b) is —OH and R^(5d) is —C₁₋₄alkyl, for example, R⁵ canbe —CH₂—N(OH)C(O)CH₃. In another embodiment, R^(5b) is H and R^(5d) is—CH₂SH, for example, R⁵ can be —C₀₋₁alkylene-NHC(O)CH₂SH, for example,—NHC(O)CH₂SH or —CH₂NHC(O)—CH₂SH.

In yet another embodiment, R⁵ is —NH—C₀₋₁alkylene-P(O)(OR^(5e))₂. TheR^(5e) moiety is H, —C₁₋₆alkyl, —C₁₋₃alkylenearyl,—C₁₋₃alkyleneheteroaryl, —C₃₋₇cycloalkyl, —CH(CH₃)—O—C(O)R^(5ea),

The R^(5ea) group is —O—C₁₋₆alkyl, —O—C₃₋₇cycloalkyl, —NR^(5eb)R^(5ec),or —CH(NH₂)CH₂COOCH₃. R^(5eb) and R^(5ec) are independently selectedfrom H, —C₁₋₄alkyl, and —C₁₋₃alkylenearyl (for example, benzyl). R^(5eb)and R^(5ec) may also be taken together to form —(CH₂)₃₋₆—. In oneembodiment, R^(5e) is H, for example, R⁵ can be —NH—CH₂—P(O)(OH)₂.

In one embodiment, R⁵ is —C₀₋₃alkylene-P(O)OR^(5e)R^(5f). The R^(5f)moiety is H, —C₁₋₄alkyl, —C₀₋₃alkylenearyl,—C₁₋₃alkylene-NR^(5fa)R^(5fb), or—C₁₋₃alkylene(aryl)-C₀₋₃alkylene-NR^(5fa)R^(5fb). The R^(5fa) andR^(5fb) groups are independently selected from H and —C₁₋₄alkyl. In oneembodiment, R^(5e) is H, for example, R⁵ can be—C₀₋₃alkylene-P(O)(OH)R^(5f).

In one embodiment, R⁵ is —C₀₋₂alkylene-CHR^(5g)—COOH. The R^(5g) moietyis H, C₁₋₆alkyl, —C₁₋₃alkylenearyl, or —CH₂—O—(CH₂)₂—OCH₃. In oneembodiment, R^(5g) is —CH₂—O—(CH₂)₂—OCH₃, for example R⁵ can be—CH₂—CH—[CH₂—O—(CH₂)₂—OCH₃]—COOH. In another embodiment, R^(5g) is H,for example R⁵ can be —CH₂COOH.

In one embodiment, R⁵ is —C₀₋₃alkylene-C(O)NR^(5h)—CHR^(5i)—COOH. TheR^(5h) moiety is H or —C₁₋₄alkyl. The R^(5i) moiety is H, —C₁₋₄alkyl, or—C₀₋₃alkylenearyl. In one embodiment, R^(5h) is H and R^(5i) is—C₀₋₃alkylenearyl, and the aryl is optionally substituted with 1 to 3substituents such as —OH, for example, R⁵ can be—C(O)NH—CH(CH₂-phenyl-OH)(COOH).

In another embodiment, R⁵ is —C₀₋₃alkylene-S—SR^(5j), and the R^(5j)moiety is —C₁₋₆alkyl, aryl, or —CH₂CH(NH₂)COOH. Examples of such R⁵groups include —C₀₋₃alkylene-S—S—CH₃, —C₀₋₃alkylene-S—S-phenyl, and—C₀₋₃alkylene-S—S—CH₂CH(NH₂)—COOH.

R⁶ is selected from —C₁₋₆alkyl, —CH₂O(CH₂)₂OCH₃,—C₁₋₆alkylene-O—C₁₋₆alkyl, —C₀₋₃alkylenearyl, —C₀₋₃alkyleneheteroaryl,and —C₀₋₃alkylene-C₃₋₇cycloalkyl. In one particular embodiment, R⁶ isselected from —C₁₋₆alkyl, —C₀₋₃alkylenearyl, and—C₀₋₃alkylene-C₃₋₇cycloalkyl. Each alkyl and each aryl in R⁶ isoptionally substituted with 1 to 7 fluoro atoms, where the term “alkyl”is intended to include divalent alkylene groups such as those present in—C₁₋₆alkylene-O—C₁₋₆alkyl and —C₀₋₃alkylene-C₃₋₇cycloalkyl, for example.In addition, each aryl and heteroaryl in R⁶ may be substituted with 1 to3-OH, —C₁₋₆alkyl, —C₂₋₄alkenyl, —C₂₋₄alkynyl, —CN, halo, —O—C₁₋₆alkyl,—S—C₁₋₆alkyl, —S(O)—C₁₋₆alkyl, —S(O)₂—C₁₋₄alkyl, -phenyl, —NO₂, —NH₂,—NH—C₁₋₆alkyl, or —N(C₁₋₆alkyl)₂ groups. Further, each of theaforementioned alkyl, alkenyl and alkynyl groups may be substituted with1 to 5 fluoro atoms.

In one embodiment, R⁶ is —C₁₋₆alkyl, for example, —CH₃, —CH₂CH₃,—CH(CH₃)₂, —(CH₂)₂CH₃, —(CH₂)₃CH₃, —CH(CH₃)CH₂CH₃, —CH₂CH(CH₃)₂,—CH₂C(CH₃)₃, —(CH₂)₂CH(CH₃)₂, and —(CH₂)₄CH₃. As noted above, each alkylin R⁶ is optionally substituted with 1 to 7 fluoro atoms. Examples ofsuch fluoro-substituted R⁶ groups include —(CH₂)₂CF₃ and —(CH₂)₃CF₃.

In another embodiment, R⁶ is —CH₂O(CH₂)₂OCH₃. In still another oneembodiment, R⁶ is —C₁₋₆alkylene-O—C₁₋₆alkyl, for example, —OCH₃ and—CH₂OCH₃.

In one embodiment, R⁶ is —C₀₋₃alkylenearyl, for example, phenyl, benzyl,—CH₂-biphenyl, —(CH₂)₂-phenyl and —CH₂-naphthalen-1-yl. The aryl may besubstituted with 1 to 3 substituents. Thus, other examples of R⁶ includemono-substituted groups such as, methylbenzyl, chlorobenzyl,fluorobenzyl, fluorophenyl, bromobenzyl, iodobenzyl, benzyl-CF₃,2-trifluoromethyl-benzyl, -benzyl-CN, and -benzyl-NO₂; anddi-substituted groups such as di-chlorobenzyl and di-fluorobenzyl. Eacharyl may also be substituted with 1 to 7 fluoro atoms. Thus, otherexamples of R⁶ include penta-fluorobenzyl.

In one particular embodiment, R⁶ is selected from —C₁₋₆alkyl and—C₀₋₃alkylenearyl (e.g., benzyl).

In one embodiment, R⁶ is —C₀₋₃alkyleneheteroaryl, for example,—CH₂-pyridyl, —CH₂-furanyl, —CH₂-thienyl, and —CH₂-thiophenyl. Inanother embodiment, R⁶ is —C₀₋₃alkylene-C₃₋₇cycloalkyl, for example,—CH₂-cyclopropyl, cyclopentyl, —CH₂-cyclopentyl, -cyclohexyl, and—CH₂-cyclohexyl.

R⁷ is H or is taken together with R⁶ to form —C₃₋₈cycloalkyl. In oneembodiment, R⁷ is H. In another embodiment, R⁷ is taken together with R⁶to form —C₃₋₈cycloalkyl, for example cyclopentyl.

One particular embodiment of the invention provides for an activecompound of formula I where Ar**—COOH represents Ar—R¹ and R⁵ is—C₀₋₃alkylene-SH. One corresponding prodrug (prodrug A) can contain athioester linkage, which can be cleaved in vivo to form the —COOH(R¹)and —C₀₋₃alkylene-SH(R⁵) moieties. Another corresponding prodrug(prodrug B, where Z is —C₁₋₆alkylene, optionally substituted with one ormore moieties such as hydroxyl, phenyl, carboxyl, and so forth),contains both an ester and a thioester group, which can be similarlycleaved in vivo, but which also releases a physiologically acceptableacid such as α-hydroxy acid (Z is —CH₂—), β-hydroxy acid (Z is—(CH₂)₂—), (R)-2-hydroxypropionic or lactic acid (Z is —CH(CH₃)—),(R)-hydroxyphenyl acetic or mandelic acid (Z is —CH(phenyl)-), salicylicacid (Z is -phenylene-), 2,3-dihydroxysuccinic or tartaric acid (Z is—CH[CH(OH)(COOH)]—), citric acid (Z is —C[CH₂COOH]₂—), hydroxy bis- andhydroxy-tris acids, and so forth.

Yet another corresponding prodrug (prodrug C) is a dimer form of prodrugA, thus containing two thioester linkages, which can both be cleaved invivo to form two active moieties, each containing the —COOH(R¹) and—C₀₋₃alkylene-SH(R⁵) moieties.

Another embodiment of the invention provides for an active compound offormula I where R⁵ is —C₀₋₃alkylene-SH, and the prodrug (prodrug D) is adimer form of the compound:

In one particular embodiment, the compound of formula I is the speciesembodied in formula Ia, Ib, or Ic:

where Ar, r, n, R², R^(2′), R³, X, and R⁵⁻⁷ are as defined for formulaI.

In one particular embodiment, the compound of formula I is the speciesembodied in formula II:

where: Ar is an aryl group selected from:

R¹ is selected from —COOH and tetrazol-5-yl; n is 0, or n is 1 and R² is—C₁₋₆alkyl; R^(2′) is selected from H and —C(O)—C₁₋₆alkyl; R³ isselected from —C₁₋₁₀alkyl and —C₀₋₅alkylene-O—C₁₋₅alkylene-H; X is—C₁₋₂alkylene-, where one —CH₂— moiety in the alkylene is replaced witha —NR^(4a)—C(O)— or —C(O)—NR^(4a)— moiety, where R^(4a) is selected fromH, —OH, and —C₁₋₄alkyl; R⁵ is selected from —C₀₋₃alkylene-SR^(5a) and—C₀₋₃alkylene-C(O)NR^(5b)R^(5c); where R^(5a) is selected from H and—C(O)—C₁₋₆alkyl; R^(5b) is selected from H, —OH, and —OC(O)—C₁₋₆alkyl;and R^(5c) is selected from H and —C₁₋₆alkyl; and R⁶ is selected from—C₁₋₆alkyl and —C₀₋₃alkylenearyl. Any of these moieties may beoptionally substituted as set forth in the description for formula I.

In one particular embodiment, the compound of formula II is the speciesembodied in formula IIa, IIb, or IIc:

where Ar, n, R², R^(2′), R³, and R⁵⁻⁶ are as defined for formula II.

In one particular embodiment, X is —C(O)—NH—. In another aspect, thisembodiment has formula Ia, Ib, Ic, II, IIa, IIb, or IIc.

In another particular embodiment, R¹ is selected from —COOH,—NHSO₂R^(1b), —SO₂NHR^(1d), —SO₂OH, —C(O)NH—SO₂R^(1c), —P(O)(OH)₂, —CN,—O—CH(R^(1e))—COOH, tetrazol-5-yl,

where R^(1b), R^(1c), R^(1d), and R^(1e), are as defined for formula I.In one particular embodiment, R¹ is selected from —COOR^(1a),—SO₂NHR^(1d), and tetrazol-5-yl. In another embodiment, R¹ is selectedfrom —COOH, —SO₂NHC(O)—C₁₋₆alkyl, and tetrazol-5-yl. In another aspect,this embodiment has formula Ia, Ib, Ic, II, IIa, IIb, or IIc.

In one particular embodiment, R¹ is —COOR^(1a), where R^(1a) is—C₁₋₆alkyl, —C₁₋₃alkylenearyl, —C₁₋₃alkyleneheteroaryl, —C₃₋₇cycloalkyl,—CH(C₁₋₄alkyl)OC(O)R^(1aa), —C₀₋₆alkylenemorpholine,

where R^(1aa) is as defined for formula I. In one aspect of theinvention, these compounds may find particular utility as prodrugs or asintermediates in the synthetic procedures described herein. In oneparticular embodiment, R¹ is —COO—C₁₋₆alkyl. In another aspect, theseembodiments have formula Ia, Ib, Ic, II, IIa, IIb, or IIc.

In one embodiment, R⁵ is selected from —C₀₋₃alkylene-SR^(5a),—C₀₋₃alkylene-C(O)NR^(5b)R^(5c), —C₀₋₃alkylene-NR^(5b)—C(O)R^(5d),—NH—C₀₋₁alkylene-P(O)(OR^(5e))₂, C₀₋₃alkylene-P(O)OR^(5e)R^(5f),—C₀₋₂alkylene-CHR^(5g)—COOH, and —C₀₋₃alkylene-C(O)NR^(5h)—CHR^(5i)COOH;where R^(5a) is H, R^(5b) is —OH, R^(5c) is H, R^(5d) is H, R^(5e) is H;and R^(5f), R^(5g)R^(5h), R^(5i) are as defined for formula I. Moreparticularly, in one embodiment, R⁵ is selected from —C₀₋₁alkylene-SH,—C₀₋₁alkylene-C(O)—N(OH)H, and —C₀₋₃alkylene-N(OH)—C(O)H. In anotherembodiment, R⁵ is selected from —C₀₋₃alkylene-SR^(5a) and—C₀₋₃alkylene-C(O)NR^(5b)R^(5c), where R^(5a) is H; R^(5b) is —OH. Inone particular embodiment, R^(5c) is H. In another aspect, thisembodiment has formula Ia, Ib, Ic, II, IIa, IIb, or IIc.

In yet another embodiment, R⁵ is selected from —C₀₋₃alkylene-SR^(5a),—C₀₋₃alkylene-C(O)NR^(5b)R^(5c), —C₀₋₃alkylene-NR^(5b)—C(O)R^(5d),—NH—C₀₋₁alkylene-P(O)(OR^(5e))₂, —C₀₋₃alkylene-P(O)OR^(5e)R^(5f), and—C₀₋₃alkylene-S—SR^(5j); where R^(5a) is —C(O)—R^(5aa); R^(5b) is H,—OC(O)R^(5ba), —CH₂COOH, —O-benzyl, -pyridyl, or —OC(S)NR^(5bb)R^(5bc);R^(5e) is —C₁₋₆alkyl, —C₁₋₃alkylenearyl, —C₁₋₃alkyleneheteroaryl,—C₃₋₇cycloalkyl, —CH(CH₃)—O—C(O)R^(5ea),

and where R^(5aa), R^(5ba), R^(5bb), R^(5bc), R^(5c), R^(5d), R^(5ea),R^(5f), and R^(5j) are as defined for formula I. In one aspect of theinvention, these compounds may find particular utility as prodrugs or asintermediates in the synthetic procedures described herein. In anotheraspect, these embodiments have formula Ia, Ib, Ic, II, IIa, IIb, or IIc.

In one particular embodiment, R⁵ is selected from —C₀₋₃alkylene-SR^(5a)and —C₀₋₃alkylene-C(O)NR^(5b)R^(5c); where R^(5a) is selected from H and—C(O)—C₁₋₆alkyl; R^(5b) is selected from H, —OH, and —OC(O)—C₁₋₆alkyl;and R^(5c) is selected from H and —C₁₋₆alkyl. In another aspect, thisembodiment has formula Ia, Ib, Ic, II, IIa, IIb, or IIc.

In another embodiment, R¹ is selected from —COOR^(1a) where R^(1a) is H,—NHSO₂R^(1b), —SO₂NHR^(1d), —SO₂OH, —C(O)NH—SO₂R^(1c), —P(O)(OH)₂, —CN,—O—CH(R^(1e))—COOH, tetrazol-5-yl,

R⁵ is selected from —C₀₋₃alkylene-SR^(5a),—C₀₋₃alkylene-C(O)NR^(5b)R^(5c), and —C₀₋₃alkylene-NR^(5b)—C(O)R^(5d),—NH—C₀₋₁alkylene-P(O)(OR^(5e))₂, —C₀₋₃alkylene-P(O)OR^(5e)R^(5f),—C₀₋₂alkylene-CHR^(5g)—COOH, and —C₀₋₃alkylene-C(O)NR^(5h)—CHR^(5i)COOH;R^(5a) is H, R^(5b) is OH, R^(5c) is H, R^(5d) is H, R^(5e) is H; andR^(5f), R^(5g), R^(5h), R^(5i) are as defined for formula I. In oneparticular embodiment, R¹ is selected from —COOH, —SO₂NHR^(1d), andtetrazol-5-yl; and R⁵ is selected from —C₀₋₃alkylene-SH, and—C₀₋₃alkylene-C(O)N(OH)H. In another aspect, these embodiments haveformula Ia, Ib, Ic, II, IIa, IIb, or IIc.

In another embodiment, R¹ is —COOR^(1a), where R^(1a) is —C₁₋₆alkyl,—C₁₋₃alkylenearyl, —C₁₋₃alkyleneheteroaryl, —C₃₋₇cycloalkyl,—CH(C₁₋₄alkyl)OC(O)R^(1aa), —C₀₋₆alkylenemorpholine,

R⁵ is selected from —C₀₋₃alkylene-SR^(5a),—C₀₋₃alkylene-C(O)NR^(5b)R^(5c), —C₀₋₃alkylene-NR^(5b)— C(O)R^(5d),—NH—C₀₋₁alkylene-P(O)(OR^(5e))₂, —C₀₋₃alkylene-P(O)OR^(5e)R^(5f), and—C₀₋₃alkylene-S—SR^(5j); where R^(5a) is —C(O)—R^(5aa); R^(5b) is H,—OC(O)R^(5ba), —CH₂COOH, —O-benzyl, -pyridyl, or —OC(S)NR^(5bb)R^(5bc);R^(5e) is —C₁₋₆alkyl, —C₁₋₃alkylenearyl, —C₁₋₃alkyleneheteroaryl,—C₃₋₇cycloalkyl, —CH(CH₃)—O—C(O)R^(5ea),

and where R^(5aa), R^(5ba), R^(5bb), R^(5bc), R^(5c), R^(5d), R^(5ea),R^(5f), and R^(5j) are as defined for formula I. In another aspect, thisembodiment has formula Ia, Ib, Ic, II, IIa, IIb, or IIc.

A particular group of compounds of formula I are those disclosed in U.S.Provisional Application No. 61/007,129, filed on Dec. 11, 2007. Thisgroup includes compounds of formula (I′):

where: r is 0, 1 or 2; Ar′ is an aryl group selected from:

R^(1′) is selected from —COOR^(1a), —NHSO₂R^(1b), —SO₂NHR^(1d), —SO₂OH,—C(O)NH—SO₂R^(1c), —P(O)(OH)₂, —CN, —OCH(R^(1e))—COOH, tetrazol-5-yl,

R^(1a) is H, —C₁₋₆alkyl, —C₁₋₃alkylenearyl, —C₁₋₃alkyleneheteroaryl,—C₃₋₇cycloalkyl, —CH(C₁₋₄alkyl)OC(O)R^(1a′), —C₀₋₆alkylenemorpholine,

R^(1a′) is —O—C₁₋₆alkyl, —O-cycloalkyl, —NR^(1a″)R^(1a′″), or—CH(NH₂)CH₂COOCH₃; R^(1a″) and R^(1a′″) are independently selected fromH, —C₁₋₆alkyl, and benzyl, or are taken together as —(CH₂)₃₋₆—; R^(1b)is R^(1c) or —NHC(O)R^(1c); R^(1c) is —C₁₋₆alkyl,—C₀₋₆alkylene-O—R^(1c′), —C₁₋₅alkylene-NR^(1c″)R^(1c′″), or—C₀₋₄alkylenearyl; R^(1c′) is H, —C₁₋₆alkyl, or —O—C₁₋₆alkyl; R^(1c″)and R^(1c′″) are independently selected from H and —C₁₋₆alkyl, or aretaken together as —(CH₂)₂—O—(CH₂)₂— or —(CH₂)₂—N[C(O)CH₃]-(CH₂)₂—;R^(1d) is H, R^(1c), —C(O)R^(1c), or —C(O)NHR^(1c); R^(1e) is —C₁₋₄alkylor aryl; n is 0, 1, 2 or 3; each R^(2′) is independently selected from—CH₂OH, halo, —NO₂, —C₁₋₆alkyl, —C₂₋₆alkenyl, —C₃₋₆cycloalkyl, —CN,—C(O)R^(2a), —C₀₋₅alkylene-OR^(2b), —C₀₋₅alkylene-NR^(2c)R^(2d),—C₀₋₃alkylenearyl, and —C₀₋₃alkyleneheteroaryl; R^(2a) is H, —C₁₋₆alkyl,—C₃₋₆cycloalkyl, —C₀₋₃alkylenearyl, OR^(2b), or —NR^(2c)R^(2d); R^(2b)is H, —C₁₋₆alkyl, —C₃₋₆cycloalkyl, or —C₀₋₁alkylenearyl; and R^(2c) andR^(2d) are independently selected from H, —C₁₋₄alkyl, and—C₀₋₁alkylenearyl; R^(2″) is selected from H and R²; R^(3′) is selectedfrom —C₁₋₁₀alkyl, —C₂₋₁₀alkenyl, —C₃₋₁₀alkynyl, —C₀₋₃alkylene-C₃₋₇cycloalkyl, —C₂₋₃alkenylene-C₃₋₇cycloalkyl,—C₂₋₃alkynylene-C₃₋₇cycloalkyl,—C₀₋₅alkylene-NR^(3a)C₀₋₅alkylene-R^(3b),—C₀₋₅alkylene-O—C₀₋₅alkylene-R^(3b),—C₁₋₅alkylene-S—C₁₋₅alkylene-R^(3b), and —C₀₋₃alkylenearyl; R^(3a) is H,—C₁₋₆alkyl, —C₃₋₆cycloalkyl, or —C₀₋₃alkylenearyl; and R^(3b) is H,—C₁₋₆alkyl, —C₃₋₆cycloalkyl, —C₂₋₄alkenyl, —C₂₋₄alkynyl, or aryl; X′ is—C₁₋₁₂alkylene-, where at least one —CH₂— moiety in the alkylene isreplaced with a —NR^(4a)—C(O)— or —C(O)—NR^(4a)— moiety, where R^(4a) isselected from H, —OH, and —C₁₋₄alkyl; R^(5′) is selected from—C₀₋₃alkylene-SR^(5a), —C₀₋₃alkylene-C(O)NR^(5e)R^(5f),—C₀₋₃alkylene-NR^(5b)—C(O)R^(5d), —NH—C₀₋₁alkylene-P(O)(OR^(5e))₂,—C₀₋₃alkylene-P(O)OR^(5e)R^(5f), —C₀₋₂alkylene-CHR^(5g)—COOH and—C₀₋₃alkylene-C(O)NR^(5h)—CHR^(5i)—COOH; R^(5a) is H or —C(O)—R^(5a′);R^(5a′) is —C₁₋₆alkyl, —C₀₋₆alkylene-C₃₋₇cycloalkyl, —C₀₋₆alkylenearyl,—C₀₋₆alkyleneheteroaryl, —C₀₋₆alkylenemorpholine,—C₀₋₆alkylenepiperazine-CH₃, —CH(NH₂)-aa where aa is an amino acid sidechain, -2-pyrrolidine, —C₀₋₆alkylene-OR^(5a″), —OC₀₋₆alkylenearyl,—C₁₋₂alkylene-OC(O)—C₁₋₆alkyl, —C₁₋₂alkylene-OC(O)—C₀₋₆alkylenearyl, or—C₁₋₂alkylene-OC(O)—OC₁₋₆alkyl; R^(5a″) is H or —C₁₋₆alkyl; R^(5b) is H,—OH, —OC(O)R^(5b′), —CH₂COOH, —O-benzyl, -pyridyl, or—OC(S)NR^(5b″)R^(5b′″); R^(5b′) is —C₁₋₆alkyl, —OCH₂-aryl, —CH₂O-aryl,or —NR^(5b″)R^(5b′″); R^(5b″) and R^(5b′″) are independently selectedfrom H and —C₁₋₄alkyl; R^(5c) is H, —C₁₋₆alkyl, or —C(O)—R^(5c′);R^(5c′) is —C₁₋₆alkyl, —C₃₋₇cycloalkyl, aryl, or heteroaryl; R^(5d) isH, —C₁₋₄alkyl, —C₀₋₃alkylenearyl, —NR^(5d′)R^(5d″), —CH₂SH, or—O—C₁₋₆alkyl; R^(5d′) and R^(5d″) are independently selected from H and—C₁₋₄alkyl; R^(5e) is H, —C₁₋₆alkyl, —C₁₋₃alkylenearyl,—C₁₋₃alkyleneheteroaryl, —C₃₋₇cycloalkyl, —CH(CH₃)OC(O)R^(5e′),

R^(5e′) is —O—C₁₋₆alkyl, —O-cycloalkyl, —NR^(5e″)R^(5e′″), or—CH(NH₂)CH₂COOCH₃; R^(5e″) and R^(5e′″) are independently selected fromH, —C₁₋₄alkyl, and —C₁₋₃alkylenearyl, or are taken together as—(CH₂)₃₋₆—; R^(5f) is H, —C₁₋₄alkyl, —C₀₋₃alkylenearyl,—C₁₋₃alkylene-NR^(5f′)R^(5f″), or—C₁₋₃alkylene(aryl)-C₀₋₃alkylene-NR^(5f′)R^(5f″); R^(5f′) and R^(5f″)are independently selected from H and —C₁₋₄alkyl; R^(5g) is H,—C₁₋₆alkyl, —C₁₋₃alkylenearyl, or —CH₂—O—(CH₂)₂—OCH₃; R^(5h) is H or—C₁₋₄alkyl; and R^(5i) is H, —C₁₋₄alkyl, or —C₀₋₃alkylenearyl; R^(6′) isselected from —C₁₋₆alkyl, —CH₂O(CH₂)₂OCH₃, —C₁₋₆alkylene-O—C₁₋₆alkyl,—C₀₋₃alkylenearyl, —C₀₋₃alkyleneheteroaryl, and—C₀₋₃alkylene-C₃₋₇cycloalkyl; and R^(7′) is H or is taken together withR⁶ to form —C₃₋₈cycloalkyl; wherein each —CH₂— group in —(CH₂)_(r)— isoptionally substituted with 1 or 2 substituents independently selectedfrom —C₁₋₄alkyl and fluoro; each carbon atom in the alkylene moiety in Xis optionally substituted with one or more R^(4b) groups and one —CH₂—moiety in X may be replaced with a group selected from—C₄₋₈cycloalkylene-, —CR^(4d)═CH—, and —CH═CR^(4d)—; wherein R^(4b) isselected from —C₀₋₅alkylene-COOR^(4c), —C₁₋₆alkyl, —C₀₋₁alkylene-CONH₂,—C₁₋₂alkylene-OH, —C₀₋₃alkylene-C₃₋₇cycloalkyl, 1H-indol-3-yl, benzyl,and hydroxybenzyl, where R^(4c) is H or —C₁₋₄alkyl; and R^(4d) isselected from —CH₂-thiophene and phenyl; each alkyl and each aryl in R¹,R², R^(2′), R³, R^(4a-4d), and R⁵⁻⁶ is optionally substituted with 1 to7 fluoro atoms; each ring in Ar and each aryl in R¹, R², R^(2′), R³, andR⁵⁻⁶ is optionally substituted with 1 to 3 substituents independentlyselected from —OH, —C₁₋₆alkyl, —C₂₋₄alkenyl, —C₂₋₄alkynyl, —CN, halo,—O—C₁₋₆alkyl, —S—C₁₋₆alkyl, —S(O)—C₁₋₆alkyl, —S(O)₂—C₁₋₄alkyl, -phenyl,—NO₂, —NH₂, —NH—C₁₋₆alkyl and —N(C₁₋₆alkyl)₂, wherein each alkyl,alkenyl and alkynyl is optionally substituted with 1 to 5 fluoro atoms;and pharmaceutically acceptable salts thereof.

In addition, particular compounds of formula I that are of interestinclude those set forth in the Examples below, as well as thepharmaceutically acceptable salts thereof.

DEFINITIONS

When describing the compounds, compositions, methods and processes ofthe invention, the following terms have the following meanings unlessotherwise indicated. Additionally, as used herein, the singular forms“a,” “an,” and “the” include the corresponding plural forms unless thecontext of use clearly dictates otherwise. The terms “comprising”,“including,” and “having” are intended to be inclusive and mean thatthere may be additional elements other than the listed elements. Allnumbers expressing quantities of ingredients, properties such asmolecular weight, reaction conditions, and so forth used herein are tobe understood as being modified in all instances by the term “about,”unless otherwise indicated. Accordingly, the numbers set forth hereinare approximations that may vary depending upon the desired propertiessought to be obtained by the present invention. At least, and not as anattempt to limit the application of the doctrine of equivalents to thescope of the claims, each number should at least be construed in lightof the reported significant digits and by applying ordinary roundingtechniques.

The term “alkyl” means a monovalent saturated hydrocarbon group whichmay be linear or branched. Unless otherwise defined, such alkyl groupstypically contain from 1 to 10 carbon atoms and include, for example,—C₁₋₄alkyl, —C₁₋₆alkyl, and —C₁₋₁₀alkyl. Representative alkyl groupsinclude, by way of example, methyl, ethyl, n-propyl, isopropyl, n-butyl,s-butyl, isobutyl, t-butyl, n-pentyl, n-hexyl, n-heptyl, n-octyl,n-nonyl, n-decyl and the like.

When a specific number of carbon atoms is intended for a particular termused herein, the number of carbon atoms is shown preceding the term assubscript. For example, the term “—C₁₋₆alkyl” means an alkyl grouphaving from 1 to 6 carbon atoms, and the term “—C₃₋₇cycloalkyl” means acycloalkyl group having from 3 to 7 carbon atoms, respectively, wherethe carbon atoms are in any acceptable configuration.

The term “alkylene” means a divalent saturated hydrocarbon group thatmay be linear or branched. Unless otherwise defined, such alkylenegroups typically contain from 0 to 12 carbon atoms and include, forexample, —C₀₋₁alkylene-, —C₀₋₂alkylene-, —C₀₋₃alkylene-, —C₀₋₅alkylene-,—C₀₋₆alkylene-, —C₁₋₂alkylene- and —C₁₋₁₂alkylene-. Representativealkylene groups include, by way of example, methylene, ethane-1,2-diyl(“ethylene”), propane-1,2-diyl, propane-1,3-diyl, butane-1,4-diyl,pentane-1,5-diyl and the like. It is understood that when the alkyleneterm include zero carbons such as —C₀₋₁alkylene- or —C₀₋₅alkylene-, suchterms are intended to include the absence of carbon atoms, that is, thealkylene group is not present except for a covalent bond attaching thegroups separated by the alkylene term.

The term “alkylthio” means a monovalent group of the formula —S-alkyl,where alkyl is as defined herein. Unless otherwise defined, suchalkylthio groups typically contain from 2 to 10 carbon atoms andinclude, for example, —S—C₁₋₄alkyl and —S—C₁₋₆alkyl. Representativealkylthio groups include, by way of example, ethylthio, propylthio,isopropylthio, butylthio, s-butylthio and t-butylthio.

The term “alkenyl” means a monovalent unsaturated hydrocarbon groupwhich may be linear or branched and which has at least one, andtypically 1, 2 or 3, carbon-carbon double bonds. Unless otherwisedefined, such alkenyl groups typically contain from 2 to 10 carbon atomsand include, for example, —C₂₋₄alkenyl and —C₂₋₁₀alkenyl. Representativealkenyl groups include, by way of example, ethenyl, n-propenyl,isopropenyl, n-but-2-enyl, n-hex-3-enyl and the like. The term“alkenylene” means a divalent alkenyl group, and includes groups such as—C₂₋₃alkenylene-.

The term “alkoxy” means a monovalent group of the formula —O-alkyl,where alkyl is as defined herein. Unless otherwise defined, such alkoxygroups typically contain from 2 to 10 carbon atoms and include, forexample, —O—C₁₋₄alkyl and —O—C₁₋₆alkyl. Representative alkoxy groupsinclude, by way of example, methoxy, ethoxy, n-propoxy, isopropoxy,n-butoxy, sec-butoxy, isobutoxy, t-butoxy and the like.

The term “alkynyl” means a monovalent unsaturated hydrocarbon groupwhich may be linear or branched and which has at least one, andtypically 1, 2 or 3, carbon-carbon triple bonds. Unless otherwisedefined, such alkynyl groups typically contain from 2 to 10 carbon atomsand include, for example, —C₂₋₄alkynyl and —C₃₋₁₀alkynyl. Representativealkynyl groups include, by way of example, ethynyl, n-propynyl,n-but-2-ynyl, n-hex-3-ynyl and the like. The term “alkynylene” means adivalent alkynyl group and includes groups such as —C₂₋₃alkynylene-.

Amino acid residues are often designated as —C(O)—CHR—NH—, where the Rmoiety is referred to as the “amino acid side chain.” Thus, for theamino acid valine, HO—C(O)—CH[—CH(CH₃)₂]—NH₂, the side chain is—CH(CH₃)₂. The term “amino acid side chain” is intended to include sidechains of the twenty common naturally occurring amino acids: alanine,arginine, asparagine, aspartic acid, cysteine, glutamic acid, glutamine,glycine, histidine, isoleucine, leucine, lysine, methionine,phenylalanine, proline, serine, threonine, tryptophan, tyrosine, andvaline. Of particular interest are the side chains of non-polar aminoacids such as isoleucine, leucine, and valine.

The term “aryl” means a monovalent aromatic hydrocarbon having a singlering (e.g., phenyl) or fused rings. Fused ring systems include thosethat are fully unsaturated (e.g., naphthalene) as well as those that arepartially unsaturated (e.g., 1,2,3,4-tetrahydronaphthalene). Unlessotherwise defined, such aryl groups typically contain from 6 to 10carbon ring atoms and include, for example, —C₆₋₁₀aryl. Representativearyl groups include, by way of example, phenyl and naphthalene-1-yl,naphthalene-2-yl, and the like. The term “arylene” means a divalent arylgroup such as phenylene.

The term “cycloalkyl” means a monovalent saturated carbocyclichydrocarbon group. Unless otherwise defined, such cycloalkyl groupstypically contain from 3 to 10 carbon atoms and include, for example,—C₃₋₅cycloalkyl, —C₃₋₆cycloalkyl and —C₃₋₇cycloalkyl. Representativecycloalkyl groups include, by way of example, cyclopropyl, cyclobutyl,cyclopentyl, cyclohexyl and the like. The term “cycloalkylene” means adivalent aryl group such as —C₄₋₈cycloalkylene.

The term “halo” means fluoro, chloro, bromo and iodo.

As used herein, the phrase “having the formula” or “having thestructure” is not intended to be limiting and is used in the same waythat the term “comprising” is commonly used.

The term “heteroaryl” means a monovalent aromatic group having a singlering or two fused rings and containing in the ring(s) at least oneheteroatom (typically 1 to 3 heteroatoms) selected from nitrogen, oxygenor sulfur. Unless otherwise defined, such heteroaryl groups typicallycontain from 5 to 10 total ring atoms and include, for example,—C₂₋₉heteroaryl. Representative heteroaryl groups include, by way ofexample, monovalent species of pyrrole, imidazole, thiazole, oxazole,furan, thiophene, triazole, pyrazole, isoxazole, isothiazole, pyridine,pyrazine, pyridazine, pyrimidine, triazine, indole, benzofuran,benzothiophene, benzoimidazole, benzthiazole, quinoline, isoquinoline,quinazoline, quinoxaline and the like, where the point of attachment isat any available carbon or nitrogen ring atom.

The term “optionally substituted” means that group in question may beunsubstituted or it may be substituted one or several times, such as 1to 3 times or 1 to 5 times. For example, an alkyl group that is“optionally substituted” with 1 to 5 fluoro atoms, may be unsubstituted,or it may contain 1, 2, 3, 4, or 5 fluoro atoms.

The term “pharmaceutically acceptable” refers to a material that is notbiologically or otherwise unacceptable when used in the invention. Forexample, the term “pharmaceutically acceptable carrier” refers to amaterial that can be incorporated into a composition and administered toa patient without causing unacceptable biological effects or interactingin an unacceptable manner with other components of the composition. Suchpharmaceutically acceptable materials typically have met the requiredstandards of toxicological and manufacturing testing, and include thosematerials identified as suitable inactive ingredients by the U.S. Foodand Drug administration.

The term “pharmaceutically acceptable salt” means a salt prepared from abase or an acid which is acceptable for administration to a patient,such as a mammal (e.g., salts having acceptable mammalian safety for agiven dosage regime). However, it is understood that the salts coveredby the invention are not required to be pharmaceutically acceptablesalts, such as salts of intermediate compounds that are not intended foradministration to a patient. Pharmaceutically acceptable salts can bederived from pharmaceutically acceptable inorganic or organic bases andfrom pharmaceutically acceptable inorganic or organic acids. Inaddition, when a compound of formula I contains both a basic moiety,such as an amine, pyridine or imidazole, and an acidic moiety such as acarboxylic acid or tetrazole, zwitterions may be formed and are includedwithin the term “salt” as used herein. Salts derived frompharmaceutically acceptable inorganic bases include ammonium, calcium,copper, ferric, ferrous, lithium, magnesium, manganic, manganous,potassium, sodium, and zinc salts, and the like. Salts derived frompharmaceutically acceptable organic bases include salts of primary,secondary and tertiary amines, including substituted amines, cyclicamines, naturally-occurring amines and the like, such as arginine,betaine, caffeine, choline, N,N′-dibenzylethylenediamine, diethylamine,2-diethylaminoethanol, 2-dimethylaminoethanol, ethanolamine,ethylenediamine, N-ethylmorpholine, N-ethylpiperidine, glucamine,glucosamine, histidine, hydrabamine, isopropylamine, lysine,methylglucamine, morpholine, piperazine, piperadine, polyamine resins,procaine, purines, theobromine, triethylamine, trimethylamine,tripropylamine, tromethamine and the like. Salts derived frompharmaceutically acceptable inorganic acids include salts of boric,carbonic, hydrohalic (hydrobromic, hydrochloric, hydrofluoric orhydroiodic), nitric, phosphoric, sulfamic and sulfuric acids. Saltsderived from pharmaceutically acceptable organic acids include salts ofaliphatic hydroxyl acids (e.g., citric, gluconic, glycolic, lactic,lactobionic, malic, and tartaric acids), aliphatic monocarboxylic acids(e.g., acetic, butyric, formic, propionic and trifluoroacetic acids),amino acids (e.g., aspartic and glutamic acids), aromatic carboxylicacids (e.g., benzoic, p-chlorobenzoic, diphenylacetic, gentisic,hippuric, and triphenylacetic acids), aromatic hydroxyl acids (e.g.,o-hydroxybenzoic, p-hydroxybenzoic, 1-hydroxynaphthalene-2-carboxylicand 3-hydroxynaphthalene-2-carboxylic acids), ascorbic, dicarboxylicacids (e.g., fumaric, maleic, oxalic and succinic acids), glucoronic,mandelic, mucic, nicotinic, orotic, pamoic, pantothenic, sulfonic acids(e.g., benzenesulfonic, camphosulfonic, edisylic, ethanesulfonic,isethionic, methanesulfonic, naphthalenesulfonic,naphthalene-1,5-disulfonic, naphthalene-2,6-disulfonic andp-toluenesulfonic acids), xinafoic acid, and the like.

As used herein, the term “prodrug” is intended to mean an inactive (orsignificantly less active) precursor of a drug that is converted intoits active form in the body under physiological conditions, e.g., bynormal metabolic processes. The term is also intended to include certainprotected derivatives of compounds of formula I that may be made priorto a final deprotection stage. Such compounds may not possesspharmacological activity at AT₁ and/or NEP, but may be administeredorally or parenterally and thereafter metabolized in the body to formcompounds of the invention which are pharmacologically active at AT₁and/or NEP. Thus, all protected derivatives and prodrugs of compoundsformula I are included within the scope of the invention. Prodrugs ofcompounds of formula I having a free carboxyl, sulfhydryl or hydroxygroup can be readily synthesized by techniques that are well known inthe art. These prodrug derivatives are then converted by solvolysis orunder physiological conditions to be the free carboxyl, sulfhydryland/or hydroxy compounds. Exemplary prodrugs include: esters includingC₁₋₆alkylesters and aryl-C₁₋₆alkylesters, carbonate esters, hemi-esters,phosphate esters, nitro esters, sulfate esters, sulfoxides, amides,carbamates, azo-compounds, phosphamides, glycosides, ethers, acetals,ketals, and disulfides. In one embodiment, the compounds of formula Ihave a free sulfhydryl or a free carboxyl and the prodrug is an esterderivative.

The term “protected derivatives thereof” means a derivative of thespecified compound in which one or more functional groups of thecompound are protected or blocked from undergoing undesired reactionswith a protecting or blocking group. Functional groups that may beprotected include, by way of example, carboxy groups, amino groups,hydroxyl groups, thiol groups, carbonyl groups and the like.Representative protecting groups for carboxy groups include esters (suchas a p-methoxybenzyl ester), amides and hydrazides; for amino groups,carbamates (such as t-butoxycarbonyl) and amides; for hydroxyl groups,ethers and esters; for thiol groups, thioethers and thioesters; forcarbonyl groups, acetals and ketals; and the like. Such protectinggroups are well-known to those skilled in the art and are described, forexample, in T. W. Greene and G. M. Wuts, Protective Groups in OrganicSynthesis, Third Edition, Wiley, New York, 1999, and references citedtherein.

The term “solvate” means a complex or aggregate formed by one or moremolecules of a solute, e.g., a compound of formula I or apharmaceutically acceptable salt thereof, and one or more molecules of asolvent. Such solvates are typically crystalline solids having asubstantially fixed molar ratio of solute and solvent. Representativesolvents include, by way of example, water, methanol, ethanol,isopropanol, acetic acid and the like. When the solvent is water, thesolvate formed is a hydrate.

The term “therapeutically effective amount” means an amount sufficientto effect treatment when administered to a patient in need thereof,i.e., the amount of drug needed to obtain the desired therapeuticeffect. For example, a therapeutically effective amount for treatinghypertension is an amount of compound needed to, for example, reduce,suppress, eliminate or prevent the symptoms of hypertension, or to treatthe underlying cause of hypertension. In one embodiment, atherapeutically effective amount is that amount needed to reduce bloodpressure or the amount needed to maintain normal blood pressure. On theother hand, the term “effective amount” means an amount sufficient toobtain a desired result, which may not necessary be a therapeuticresult. For example, when studying a system comprising an AT₁ receptor,an “effective amount” may be the amount needed to antagonize thereceptor.

The term “treating” or “treatment” as used herein means the treating ortreatment of a disease or medical condition (such as hypertension) in apatient, such as a mammal (particularly a human) that includes one ormore of the following: (a) preventing the disease or medical conditionfrom occurring, such as by prophylactic treatment of a patient; (b)ameliorating the disease or medical condition, such as by eliminating orcausing regression of the disease or medical condition in a patient; (c)suppressing the disease or medical condition, such as by slowing orarresting the development of the disease or medical condition in apatient; or (d) alleviating the symptoms of the disease or medicalcondition in a patient. For example, the term “treating hypertension”would include preventing hypertension from occurring, amelioratinghypertension, suppressing hypertension, and alleviating the symptoms ofhypertension (e.g., lowering blood pressure). The term “patient” isintended to include those mammals, such as humans, that are in need oftreatment or disease prevention, that are presently being treated fordisease prevention or treatment of a specific disease or medicalcondition, as well as test subjects in which compounds of the inventionare being evaluated or being used in a assay, for example an animalmodel.

All other terms used herein are intended to have their ordinary meaningas understood by those of ordinary skill in the art to which theypertain.

General Synthetic Procedures

Compounds of the invention can be prepared from readily availablestarting materials using the following general methods, the proceduresset forth in the Examples, or by using other methods, reagents, andstarting materials that are known to those of ordinary skill in the art.Although the following procedures may illustrate a particular embodimentof the invention, it is understood that other embodiments of theinvention can be similarly prepared using the same or similar methods orby using other methods, reagents and starting materials known to thoseof ordinary skill in the art. It will also be appreciated that wheretypical or preferred process conditions (i.e., reaction temperatures,times, mole ratios of reactants, solvents, pressures, etc.) are given,other process conditions can also be used unless otherwise stated. Whileoptimum reaction conditions will typically vary depending on variousreaction parameters such as the particular reactants, solvents andquantities used, those of ordinary skill in the art can readilydetermine suitable reaction conditions using routine optimizationprocedures.

Additionally, as will be apparent to those skilled in the art,conventional protecting groups may be necessary or desired to preventcertain functional groups from undergoing undesired reactions. Thechoice of a suitable protecting group for a particular functional groupas well as suitable conditions and reagents for protection anddeprotection of such functional groups are well-known in the art.Protecting groups other than those illustrated in the proceduresdescribed herein may be used, if desired. For example, numerousprotecting groups, and their introduction and removal, are described inT. W. Greene and G. M. Wuts, Protective Groups in Organic Synthesis,supra. More specifically, the following abbreviations and reagents areused in the schemes presented below:

P¹ represents an “amino-protecting group,” a term used herein to mean aprotecting group suitable for preventing undesired reactions at an aminogroup. Representative amino-protecting groups include, but are notlimited to, t-butoxycarbonyl (BOC), trityl (Tr), benzyloxycarbonyl(Cbz), 9-fluorenylmethoxycarbonyl (Fmoc), formyl, trimethylsilyl (TMS),t-butyldimethylsilyl (TBDMS), and the like. Standard deprotectiontechniques are used to remove the P¹ group. For example, an N—BOC groupcan be removed using an acidic reagent such as TFA in DCM or HCl in1,4-dioxane, while a Cbz group can be removed by employing catalytichydrogenation conditions such as H₂ (1 atm) and 10% Pd/C in an alcoholicsolvent (“H₂/Pd/C”).

P² represents a “carboxy-protecting group,” a term used herein to mean aprotecting group suitable for preventing undesired reactions at acarboxy group. Representative carboxy-protecting groups include, but arenot limited to, methyl, ethyl, t-butyl, benzyl (Bn), p-methoxybenzyl(PMB), 9-fluoroenylmethyl (Fm), trimethylsilyl (TMS),t-butyldimethylsilyl (TBDMS), diphenylmethyl (benzhydryl, DPM) and thelike. Standard deprotection techniques and reagents are used to removethe P² group, and may vary depending upon which group is used. Forexample, NaOH is commonly used when P² is methyl, an acid such as TFA orHCl is commonly used when P² is t-butyl, and H₂/Pd/C may be used when P²is benzyl.

P³ represents a “thiol-protecting group,” a term used herein to mean aprotecting group suitable for preventing undesired reactions at a thiolgroup. Representative thiol-protecting groups include, but are notlimited to, ethers, esters such as —C(O)CH₃, and the like. Standarddeprotection techniques and reagents such as NaOH, primary alkylamines,and hydrazine, may be used to remove the P³ group.

P⁴ represents a “tetrazole-protecting group,” a term used herein to meana protecting group suitable for preventing undesired reactions at atetrazole group. Representative tetrazole-protecting groups include, butare not limited to trityl, benzoyl, and diphenylmethyl. Standarddeprotection techniques and reagents such as TFA in DCM or HCl in1,4-dioxane are used to remove the P⁴ group.

P⁵ represents a “hydroxyl-protecting group,” a term used herein to meana protecting group suitable for preventing undesired reactions at ahydroxyl group. Representative hydroxyl-protecting groups include, butare not limited to C₁₋₆alkyls, silyl groups including triC₁₋₆alkylsilylgroups, such as trimethylsilyl (TMS), triethylsilyl (TES), andtert-butyldimethylsilyl (TBDMS); esters (acyl groups) includingC₁₋₆alkanoyl groups, such as formyl, acetyl, and pivaloyl, and aromaticacyl groups such as benzoyl; arylmethyl groups such as benzyl (Bn),p-methoxybenzyl (PMB), 9-fluorenylmethyl (Fm), and diphenylmethyl(benzhydryl, DPM); and the like. Standard deprotection techniques andreagents are used to remove the P⁵ group, and may vary depending uponwhich group is used. For example, H₂/Pd/C is commonly used when P⁵ isbenzyl, while NaOH is commonly used when P⁵ is an acyl group.

P⁶ represents a “sulfonamide-protecting group,” a term used herein tomean a protecting group suitable for preventing undesired reactions at asulfonamide group. Representative sulfonamide-protecting groups include,but are not limited to t-butyl and acyl groups. Exemplary acyl groupsinclude aliphatic lower acyl groups such as the formyl, acetyl,phenylacetyl, butyryl, isobutyryl, valeryl, isovaleryl and pivaloylgroups, and aromatic acyl groups such as the benzoyl and4-acetoxybenzoyl. Standard deprotection techniques and reagents are usedto remove the P⁶ group, and may vary depending upon which group is used.For example, HCl is commonly used when P⁶ is t-butyl, while NaOH iscommonly used when P⁶ is an acyl group.

P⁷ represents a “phosphate -protecting group or phosphinate-protectinggroup,” a term used herein to mean a protecting group suitable forpreventing undesired reactions at a phosphate or phosphinate group.Representative phosphate and phosphinate protecting groups include, butare not limited to C₁₋₄alkyls, aryl (e.g., phenyl) and substituted aryls(e.g., chlorophenyl and methylphenyl). The protected group can berepresented by —P(O)(OR)₂, where R is a group such as a C₁₋₆alkyl orphenyl. Standard deprotection techniques and reagents such asTMS-I/2,6-lutidine, and H₂/Pd/C are used to remove the P⁷ group such asethyl, and benzyl, respectively.

In addition, L is used to designate a “leaving group,” a term usedherein to mean a functional group or atom which can be displaced byanother functional group or atom in a substitution reaction, such as anucleophilic substitution reaction. By way of example, representativeleaving groups include chloro, bromo and iodo groups; sulfonic estergroups, such as mesylate, triflate, tosylate, brosylate, nosylate andthe like; and acyloxy groups, such as acetoxy, trifluoroacetoxy and thelike.

Suitable bases for use in these schemes include, by way of illustrationand not limitation, potassium carbonate, calcium carbonate, sodiumcarbonate, triethylamine, pyridine, 1,8-diazabicyclo-[5.4.0]undec-7-ene(DBU), N,N-diisopropylethylamine (DIPEA), sodium hydroxide, potassiumhydroxide, potassium t-butoxide, and metal hydrides.

Suitable inert diluents or solvents for use in these schemes include, byway of illustration and not limitation, tetrahydrofuran (THF),acetonitrile (MeCN), N,N-dimethylformamide (DMF), dimethyl sulfoxide(DMSO), toluene, dichloromethane (DCM), chloroform (CHCl₃), carbontetrachloride (CCl₄), 1,4-dioxane, methanol, ethanol, water, and thelike.

Suitable carboxylic acid/amine coupling reagents includebenzotriazol-1-yloxytris(dimethylamino)phosphonium hexafluorophosphate(BOP), benzotriazol-1-yloxytripyrrolidinophosphonium hexafluorophosphate(PyBOP), O-(7-azabenzotriazol-1-yl-N,N,N′,N′ tetramethyluroniumhexafluorophosphate (HATU), dicyclohexylcarbodiimide (DCC),N-(3-dimethylaminopropyl)-N′-ethylcarbodiimide (EDC),carbonyldiimidazole (CDI), and the like. Coupling reactions areconducted in an inert diluent in the presence of a base, and areperformed under conventional amide bond-forming conditions.

All reactions are typically conducted at a temperature within the rangeof about −78° C. to 100° C., for example at room temperature. Reactionsmay be monitored by use of thin layer chromatography (TLC), highperformance liquid chromatography (HPLC), and/or LCMS until completion.Reactions may be complete in minutes, or may take hours, typically from1-2 hours and up to 48 hours. Upon completion, the resulting mixture orreaction product may be further treated in order to obtain the desiredproduct. For example, the resulting mixture or reaction product may besubjected to one or more of the following procedures: concentrating orpartitioning (e.g., between EtOAc and water or between 5% THF in EtOAcand IM phosphoric acid); extraction (e.g., with EtOAc, CHCl₃, DCM,chloroform); washing (e.g., with saturated aqueous NaCl, saturatedNaHCO₃, Na₂CO₃ (5%), CHCl₃ or 1M NaOH); drying (e.g., over MgSO₄, overNa₂SO₄, or in vacuo); filtering; crystallizing (e.g., from EtOAc andhexane); being concentrated (e.g., in vacuo); and/or purification (e.g.,silica gel chromatography, flash chromatography, preparative HPLC,reverse phase-HPLC, or crystallization).

By way of illustration, compounds of formula I, as well as their salts,solvates, and prodrugs can be prepared by one or more of the followingexemplary processes. One method of preparing compounds of the inventioninvolves coupling compound (1) and (2), following by reacting withcompound (4), with an optional deprotection step when R¹* is a protectedform of R¹ and/or R⁵* is a protected form of R⁵, as depicted in Scheme I(R² is typically a moiety such as —CH₃ and R^(2′) is typically H):

Another method of preparing compounds of the invention involves couplingcompound (6) and (7), then reacting the product with compound (9), withan optional deprotection step, as depicted in Scheme TI (n is typically0 and R^(2′) is typically H).

The X moiety contains one or more amide groups, and therefore thecompounds of the invention may be formed by a coupling reaction underconventional amide bond-forming conditions, followed by a deprotectionstep if needed. In Schemes I and II, the A and B moieties couple to formX, and the sum of a and b is in the range of 0 to 11. Thus, one moietycomprises an amine group and one moiety comprises a carboxylic acidgroup, i.e., A is —(CH₂)_(a)—NH₂ and B is —(CH₂)_(b)—COOH or A is—(CH₂)_(a)—COOH and B is —(CH₂)_(b)—NH₂. For example, to synthesize acompound of formula I where X is —CONH—, A would be —COOH and B would be—NH₂. Similarly, A as —NH₂ and B as —COOH would couple to form —NHCO— asthe X moiety. A and B can be readily modified if a longer X is desired,whether it contains an alkylene portion or additional amide groups. Forexample, A as —CH₂NH₂ and B as —COOH would couple to form —CH₂NHCO— asthe X moiety.

It is understood that the carbon atoms in the —(CH₂)_(a) and —(CH₂)_(b)groups make up the “X” linker. Therefore, these carbon atoms may besubstituted with one or more R^(4b) groups. Furthermore, one —CH₂— groupin the —(CH₂)a or the —(CH₂)_(b) group may be replaced with a—C₄₋₈cycloalkylene-, —CR^(4d)═CH—, or —CH═CR^(4d)— group.

Ar* represents Ar—R¹*, where R¹* may represent R¹ as defined herein, ora protected form of R¹ (e.g., -tetrazol-5-yl-P⁴ or —C(O)O—P² such as—C(O)O—C₁₋₆alkyl), or a precursor of R¹ (e.g., —CN that is thenconverted to tetrazole, or nitro that is then converted to amino fromwhich the desired R¹ is prepared). R⁵* represents R⁵ as defined herein,or a protected form of R⁵. Therefore, when R¹* represents R¹ and R⁵*represents R⁵, the reaction is complete after the coupling step.

On the other hand, when R¹* represents a protected form of R¹ and/or R⁵*represents a protected form of R⁵, a subsequent global or sequentialdeprotection step would yield the non-protected compound. Similarly,when R¹* represents a precursor of R¹, a subsequent conversion stepwould yield the desired compound. Reagents and conditions for thedeprotection vary with the nature of protecting groups in the compound.Typical deprotection conditions when R⁵* represents C₀₋₃alkylene-S—P³,include treating the compound with NaOH in an alcoholic solvent at 0° C.or room temperature to yield the non-protected compound. Typicaldeprotection conditions when R¹* represents C(O)O—P² where P² refers tot-butyl include treating the compound with TFA in DCM at roomtemperature to yield the non-protected compound. Thus, one method ofpreparing compounds of the invention involves coupling compounds (1) and(2), with an optional deprotection step when R¹* is a protected form ofR¹ and/or R⁵ is a protected form of R⁵, thus forming a compound offormula I or a pharmaceutically acceptable salt thereof.

Examples of compound (1) include the commercially available7-methyl-2-propyl-3H-benzoimidazole-5-carboxylic acid. Examples ofcompound (2) include (R)-3-amino-N-benzyloxy-4-phenylbutyramide.Examples of compound (4) include 4-bromomethylbenzoic acid methyl esterand 5-(4′-bromomethylbiphenyl-2-yl)-1-trityl-1H-tetrazole. Examples ofcompound (6) include the commercially available2-ethoxy-3-[2′-(1H-tetrazol-5-yl)-biphenyl-4-ylmethyl]-3H-benzimidazole-4-carboxylicacid. Examples of compound (7) include 1-chloromethyl-3-methylbutylaminehydrochloride. Compound 9 is a salt form of the R⁵ or R⁵* substituent,for example, potassium thioacetate. Starting materials and reagents arecommercially available or can be readily synthesized by techniques knownin the art, as well as by the methods described below and in theExamples.

Preparation of Compound (2)

Compound (2) is readily synthesized by following techniques described inthe literature, for example, Neustadt et al. (1994) J. Med. Chem.37:2461-2476 and Moree et al. (1995) J. Org. Chem. 60: 5157-69, as wellas by using the exemplary procedures described below. Examples ofcompound (2), depicted without chirality, include:

Since compound (2) has a chiral center, it may be desirable tosynthesize a particular stereoisomer, and examples are as follows.

Preparation of Chiral Amino Hydroxamate Compound (2^(i))

A base such as DIPEA and a coupling agent such as EDC are added to asolution of compound (2a) in DMF containing HOBt and hydroxylaminehydrochloride. The mixture is stirred at room temperature until thereaction is complete, then concentrated in vacuo. The resulting materialis distributed between 5% THF in EtOAc and 1M phosphoric acid. Theorganic layer is collected and washed with a base such as 1M NaOH. Thealkaline aqueous layer is then acidified (e.g., with 1M phosphoricacid), and extracted with EtOAc. The organic layer is evaporated and theresidue purified by silica gel chromatography to afford compound(2^(i)). Examples of compound (2a) include(R)-3-t-butoxycarbonylamino-4-phenylbutyric acid.

Preparation of Sulfanyl Acid Compound (2^(ii))

Compound (2b) is mixed with diethylamine and cooled in an ice bath. Anaqueous formaldehyde solution (37%) is then added, and the mixturestirred at 0° C. for ˜2 hours, warmed to room temperature and stirredovernight. The mixture is then extracted with ether, washed, dried, andevaporated to dryness, to provide compound (2c). Compound (2c) is thendissolved in 1,4-dioxane, and a 1M NaOH solution is added. The mixtureis stirred at room temperature until the reaction is complete. Theorganic solvent is removed in vacuo, and the aqueous residue is rinsedwith EtOAc and acidified to pH˜1 with concentrated HCl. The product isextracted with EtOAc, dried, and evaporated to dryness to yield compound(2d). Compound (2d) is combined with thiolacetic acid (10 mL), and themixture was stirred at 80° C. until the reaction is complete, thenconcentrated to dryness to yield Compound (2^(ii)), which is dissolvedin toluene and concentrated to remove any trace of thiolacetic acid.Examples of (2b) include 2-benzylmalonic acid monoethyl ester(R⁶=benzyl) and 2-isobutylmalonic acid monoethyl ester (R⁶=isobutyl).

Preparation of Chiral Amino Sulfhydryl Dimer Compound (2^(iii))

Diisopropyl azodicarboxylate is added to a solution oftriphenylphosphine in a solvent such as THF, cooled in an ice bath. Thesolution is stirred and compound (2e) and thioacetic acid are added. Themixture is first stirred at 0° C., then at room temperature until thereaction is complete. The mixture is stripped, diluted with EtOAc, andwashed. The organic layer is dried and the filtrate evaporated todryness. The resulting material is flash chromatographed to providecompound (2f). Compound (2f) is dissolved in a suitable solvent,followed by the addition of a base such as 1M LiOH. Air is bubbledthrough the solution for 1 hour followed by the addition of solvent. Themixture is stirred at room temperature until the reaction is complete.The solution is then acidified to pH5, for example with acetic acid. Theprecipitate is filtered and rinsed producing compound (2g) as a solid,which is suspended in MeCN, then concentrated in vacuo. The recoveredmaterial is dissolved in 4M HCl in 1,4-dioxane and stirred at roomtemperature until the reaction is complete. The mixture is thenconcentrated under reduced pressure, and triturated with EtOAc. Theproduct is filtered, washed, and dried in vacuo to provide compound(2^(iii)). Examples of compound (2e) include((R)-1-benzyl-2-hydroxyethyl)carbamic acid t-butyl ester.

Preparation of Chiral Sulfanyl Acid Compound (2^(iv))

Compound (2h) is formed by dissolving a compound such as D-leucine (forR⁶=isobutyl, for example) in 3M HBr (aqueous) and cooled to 0° C. Asolution of sodium nitrite in water is added, and the mixture is stirredat 0° C. until the reaction is complete (˜2.5 hours). The mixture isthen extracted with EtOAc, washed, dried, filtered, and concentrated toafford compound (2h). Compound (2h) is combined with potassiumthioacetate and DMF, and the mixture stirred at room temperature untilthe reaction is complete. Water is added, and the mixture is thenextracted, washed, dried, filtered, and concentrated to provide compound(2^(iv)). The product is purified by silica gel chromatography. Examplesof compound (2h) include (R)-2-bromo-4-methylpentanoic acid. Examples ofcompound (2^(iv)) include (S)-2-acetylsulfanyl-4-methylpentanoic acid.

Preparation of Chiral Sulfanyl Acid Compound (2^(v))

Compound (2i), (S)-4-benzyl-2-oxazolidinone, is typically commerciallyavailable. Compound (2j) is also typically commercially available or canbe readily synthesized. For example, R⁶—CH₂—COOH (e.g., isocaproic acidor 3-phenylpropionic acid) is dissolved in methylene chloride andthionyl chloride is added. The mixture is stirred at room temperatureuntil the reaction is complete, and then concentrated to providecompound (2j). Examples of compound (2j) include 4-methylpentanoylchloride and 3-phenylpropionyl chloride.

Compound (2i) is dissolved in a suitable solvent and cooled (−78° C.)under nitrogen. n-Butyllithium in hexane is added dropwise and stirred,followed by the addition of compound (2j) dropwise. The mixture isstirred at −78° C., then warmed to 0° C. Saturated NaHCO₃ is added andthe mixture warmed to room temperature. The mixture is extracted,washed, dried, filtered and concentrated to afford compound (2k).Compound (2k) is dissolved in DCM and stirred at 0° C. under nitrogen.1M Titanium tetrachloride is added, followed by 1,3,5-trioxane, all inappropriate solvents. A second equivalent of 1M titanium tetrachlorideis added and the mixture stirred at 0° C. until the reaction iscomplete. The mixture is then quenched with saturated ammonium chloride.Appropriate solvents are added, the aqueous phase is extracted, and theorganic layers are combined, dried, filtered and concentrated to provide(2l), which is then purified by silica gel chromatography or used in thenext step without further purification. Compound (2l) is dissolved in asolvent, to which is added 9 M hydrogen peroxide in water, followed bythe dropwise addition of 1.5 M lithium hydroxide monohydrate in water.The mixture is warmed to room temperature and stirred. Optionally,potassium hydroxide may be added and the mixture heated at 60° C. thencooled at room temperature. To this is added an aqueous solution ofsodium sulfite followed by water and chloroform. The aqueous layer isextracted, acidified and extracted again. The organic layer is washed,dried, filtered, and rotovaped to provide (2m). Triphenylphosphine isdissolved in an appropriate solvent and cooled at 0° C. (ice bath).Diisopropyl azodicarboxylate is added dropwise and the mixture stirred.Compound (2m) and thioacetic acid, dissolved in an appropriate solvent,are added dropwise to the mixture. After the addition, the mixture isremoved from the ice bath and stirred at room temperature until thereaction is complete (˜3.5 hours), concentrated, and then partitioned.The organic layer is extracted and the combined aqueous extracts washed,acidified and extracted. The organic layer is washed again, dried,filtered, and rotovaped to provide compound (2V). Examples of compound(2V) include (S)-2-acetylsulfanylmethyl-4-methylpentanoic acid.

Preparation of Compound (4)

The starting material (4a) can be prepared using synthetic methods thatare reported in the literature, for example Duncia et al. (1991) J. Org.Chem. 56: 2395-400, and references cited therein. Alternatively, thestarting material in protected form (4b) may be commercially available.Using a commercially available non-protected starting material (4a), theR¹ group is first protected to form protected intermediate (4b), thenthe leaving group (L) is added to form compound (4), for example, by ahalogenation reaction. For example, a bromination reaction of a methylgroup of N-triphenylmethyl-5-[4′-methylbiphenyl-2-yl]tetrazole isdescribed in Chao et al (2005) J. Chinese Chem. Soc. 52:539-544. Inaddition, when Ar* has a —CN group, it can be subsequently converted tothe desired tetrazolyl group, which may be protected. Conversion of thenitrile group is readily achieved by reaction with a suitable azide suchas sodium azide, trialkyltin azide (particularly tributyltin azide) ortriaryltin azide. Compound (4) when Ar has one of the remaining formulasis readily synthesized using similar techniques or other methods as arewell known in the art.

Exemplary methods of preparing compound (4) include the following. Asolution of the starting material (4a) and thionyl chloride are stirredat room temperature. After completion, the mixture is concentrated invacuo to afford a solid, which is dissolved in an appropriate solventand cooled (˜0° C.). Potassium t-butoxide is then added. Uponcompletion, the mixture is partitioned, the organic layer washed, dried,filtered, and concentrated to afford compound (4b). Alternately, HCl isadded to a solution of starting material (4a) and a solvent such asmethanol. The mixture is heated to reflux, stirred until completion (˜48hours), then cooled and concentrated. The recovered material is dried invacuo to obtain intermediate (4b). Intermediate (4b), benzoyl peroxide,and N-bromosuccinimide, are dissolved in CCl₄ or benzene, and heated toreflux. The mixture is stirred until the reaction is complete, cooled toroom temperature, filtered, and concentrated in vacuo. The resultingresidue is crystallized from diethyl ether and hexane or flashchromatographed to give compound (4).

Examples of (4a) include 4′-methylbiphenyl-2-carboxylic acid,2-fluoro-4-methylbenzoic acid, and 2,3-difluoro-4-methyl-benzoic acid.Examples of (4b) includeN-triphenylmethyl-5-[4′-methylbiphenyl-2-yl]tetrazole.

Compound (4) where R¹ is —SO₂NHR^(1d) may be synthesized as follows:

The starting material, 2-bromobenzene-1-sulfonamide, is commerciallyavailable. Reaction of 2-bromobenzene-1-sulfonamide in a solvent such asDMF, with 1,1-dimethoxy-N,N-dimethylmethanamine, followed by theaddition of sodium hydrogen sulfate in water, yields2-bromo-N-[1-dimethylaminometh-(E)-ylidene]benzenesulfonamide. Thiscompound is reacted with 4-methylphenylboronic acid to yield4′-methylbiphenyl-2-sulfonic acid 1-dimethylaminometh-(E)-ylideneamide,then the —(CH₂)_(r)-L moiety is added, for example, by a halogenationreaction, to form compound (4).

Compound (4) where the Ar moiety is substituted may be synthesized asfollows:

The starting material, 2-bromobenzoic acid, is commercially available.Reaction of 2-bromobenzoic acid in a suitable solvent, with t-butylalcohol, DCC and DMAP, yields 2-bromo-benzoic acid t-butyl ester. Thiscompound is reacted with 3-fluoro-4-methylphenylboronic acid to yield3′-fluoro-4′-methyl-biphenyl-2-carboxylic acid t-butyl ester, then the—(CH₂)_(r)-L moiety is added, for example, by a halogenation reaction,to form compound (4).

Examples of (4) include 4′-bromomethylbiphenyl-2-carboxylic acid t-butylester, 4-bromomethyl-2-fluorobenzoic acid methyl ester,5-(4′-bromomethylbiphenyl-2-yl)-1-trityl-1H-tetrazole,4-bromomethylbenzoic acid methyl ester; and4-bomomethyl-2,3-difluorobenzoic acid methyl ester;4′-formyl-biphenyl-2-sulfonic acid t-butylamide;4′-aminomethylbiphenyl-2-carboxylic acid t-butyl ester; and4′-bromomethyl-3′-fluorobiphenyl-2-carboxylic acid t-butyl ester.

If desired, pharmaceutically acceptable salts of the compounds offormula I can be prepared by contacting the free acid or base form of acompound of formula I with a pharmaceutically acceptable base or acid.

Certain intermediates described herein are believed to be novel andaccordingly, such compounds are provided as further aspects of theinvention including, for example, the compounds of formulas III, IV, andV, and salts thereof:

where Ar* is Ar—R¹*; Ar, r, n, R², R^(2′), R³, X, and R⁵⁻⁷ are asdefined for formula I; and R¹* is selected from —C(O)O—P², —SO₂O—P⁵,—SO₂NH—P⁶, —P(O)(O—P⁷)₂, —OCH(CH₃)—C(O)O—P², —OCH(aryl)-C(O)O—P², andtetrazol-5-yl-P⁴; where P² is a carboxy-protecting group, P⁴ is atetrazole-protecting group, P⁵ is a hydroxyl-protecting group, P⁶ is asulfonamide-protecting group, and P⁷ is a phosphate-protecting group orphosphinate-protecting group;

where Ar, r, n, R², R^(2′), R³, X, and R⁶⁻⁷ are as defined for formulaI; R⁵* is selected from —C₀₋₃alkylene-S—P³, —C₀₋₃alkylene-C(O)NH(O—P⁵),—C₀₋₃alkylene-N(O—P⁵)—C(O)R^(5d), —C₀₋₁alkylene-NHC(O)CH₂S—P³,—NH—C₀₋₁alkylene-P(O)(O—P⁷)₂, —C₀₋₃alkylene-P(O)(O—P⁷)—R^(5f),—C₀₋₂alkylene-CHR^(5g)—C(O)O—P² and—C₀₋₃alkylene-C(O)NR^(5h)CHR^(5i)—C(O)O—P², and —C₀₋₃alkylene-S—S—P³;and R^(5d-i) are as defined for formula I; where P² is acarboxy-protecting group, P³ is a thiol-protecting group, P⁵ is ahydroxyl-protecting group, and P⁷ is a phosphate-protecting group orphosphinate-protecting group; and

where Ar* is Ar—R¹*; Ar, r, n, R², R^(2′), R³, X, and R⁶⁻⁷ are asdefined for formula I; R¹* is selected from —C(O)O—P², —SO₂O—P⁵,—SO₂NH—P⁶, —P(O)(O—P⁷)₂, —OCH(CH₃)—C(O)O—P², —OCH(aryl)-C(O)O—P², andtetrazol-5-yl-P⁴; R⁵* is selected from —C₀₋₃alkylene-S—P³,—C₀₋₃alkylene-C(O)NH(O—P⁵), —C₀₋₃alkylene-N(O—P⁵)—C(O)R^(5d),—C₀₋₁alkylene-NHC(O)CH₂S—P³, —NH—C₀₋₁ alkylene-P(O)(O—P⁷)₂,—C₀₋₃alkylene-P(O)(O—P⁷)—R^(5f), —C₀₋₂alkylene-CHR^(5g)—C(O)O—P² and—C₀₋₃alkylene-C(O)NR^(5h)CHR^(5i)—C(O)O—P², and —C₀₋₃alkylene-S—S—P³;and R^(5d-i) are as defined for formula I; where P² is acarboxy-protecting group, P³ is a thiol-protecting group, P⁴ is atetrazole-protecting group, P⁵ is a hydroxyl-protecting group, P⁶ is asulfonamide-protecting group, and P⁷ is a phosphate-protecting group orphosphinate-protecting group. Thus, another method of preparingcompounds of the invention involves deprotecting a compound of formulaIII, IV, or V.

Further details regarding specific reaction conditions and otherprocedures for preparing representative compounds of the invention orintermediates thereof are described in the Examples set forth below.

Utility

Compounds of the invention possess angiotensin TI type 1 (AT₁) receptorantagonist activity. In one embodiment, compounds of the invention areselective for inhibition of the AT₁ receptor over the AT₂ receptor.Compounds of the invention also possess neprilysin (NEP) inhibitionactivity, i.e., the compounds are able to inhibit enzyme-substrateactivity. In another embodiment, the compounds do not exhibitsignificant inhibitory activity at the angiotensin-converting enzyme.Compounds of formula I may be active drugs as well as prodrugs. Thus,when discussing the activity of compounds of the invention, it isunderstood that any such prodrugs have the expected AT₁ and NEP activityonce metabolized.

One measure of the affinity of a compound for the AT₁ receptor is theinhibitory constant (K_(i)) for binding to the AT₁ receptor. The pK_(i)value is the negative logarithm to base 10 of the K_(i). One measure ofthe ability of a compound to inhibit NEP activity is the inhibitoryconcentration (IC₅₀), which is the concentration of compound thatresults in half-maximal inhibition of substrate conversion by the NEPenzyme. The pIC₅₀ value is the negative logarithm to base 10 of theIC₅₀. Compounds of the invention that have both AT₁receptor-antagonizing activity and NEP enzyme-inhibiting activity are ofparticular interest, including those that exhibit a pK_(i) at the AT₁receptor greater than or equal to about 5.0, and exhibit a pIC₅₀ for NEPgreater than or equal to about 5.0.

In one embodiment, compounds of interest have a pK_(i) at the AT₁receptor ≧about 6.0, a pK_(i) at the AT₁ receptor ≧about 7.0, or apK_(i) at the AT₁ receptor ≧about 8.0. Compounds of interest alsoinclude those having a pIC₅₀ for NEP ≧about 6.0 or a pIC₅₀ for NEP≧about 7.0. In another embodiment, compounds of interest have a pK_(i)at the AT₁ receptor within the range of about 8.0-10.0 and a pIC₅₀ forNEP within the range of about 7.0-10.0.

In another embodiment, compounds of particular interest have a pK_(i)for binding to an AT₁ receptor greater than or equal to about 7.5 and aNEP enzyme pIC₅₀ greater than or equal to about 7.0. In anotherembodiment, compounds of interest have a pK_(i) greater than or equal toabout 8.0 and a pIC₅₀ greater than or equal to about 8.0.

It is noted that in some cases, compounds of the invention, while stillhaving dual activity, may possess either weak AT₁ receptor antagonistactivity or weak NEP inhibition activity. In such cases, those of skillin the art will recognize that these compounds still have utility asprimarily either a NEP inhibitor or an AT₁ receptor antagonist,respectively, or have utility as research tools.

Exemplary assays to determine properties of compounds of the invention,such as the AT₁ receptor binding and/or NEP inhibiting activity, aredescribed in the Examples and include by way of illustration and notlimitation, assays that measure AT₁ and AT₂ binding (described in Assay1), and NEP inhibition (described in Assay 2). Useful secondary assaysinclude assays to measure ACE inhibition (also described in Assay 2) andaminopeptidase P (APP) inhibition (described in Sulpizio et al. (2005)JPET 315:1306-1313). A pharmacodynamic assay to assess the in vivoinhibitory potencies for ACE, AT₁, and NEP in anesthetized rats isdescribed in Assay 3 (see also Seymour et al. Hypertension 7(SupplI):I-35-I-42, 1985 and Wigle et al. Can. J. Physiol. Pharmacol.70:1525-1528, 1992), where AT₁ inhibition is measured as the percentinhibition of the angiotensin II pressor response, ACE inhibition ismeasured as the percent inhibition of the angiotensin I pressorresponse, and NEP inhibition is measured as increased urinary cyclicguanosine 3′, 5′-monophosphate (cGMP) output. Useful in vivo assaysinclude the conscious spontaneously hypertensive rat (SHR) model, whichis a renin dependent hypertension model that is useful for measuring AT₁receptor blocking (described in Assay 4; see also Intengan et al. (1999)Circulation 100(22):2267-2275 and Badyal et al. (2003) Indian Journal ofPharmacology 35:349-362), and the conscious desoxycorticosteroneacetate-salt (DOCA-salt) rat model, which is a volume dependenthypertension model that is useful for measuring NEP activity (describedin Assay 5; see also Trapani et al. (1989) J. Cardiovasc. Pharmacol.14:419-424, Intengan et al. (1999) Hypertension 34(4):907-913, andBadyal et al. (2003) supra). Both the SHR and DOCA-salt models areuseful for evaluating the ability of a test compound to reduce bloodpressure. The DOCA-salt model is also useful to measure a testcompound's ability to prevent or delay a rise in blood pressure.Compounds of the invention are expected to antagonize the AT₁ receptorand/or inhibit the NEP enzyme in any of the assays described herein, orassays of a similar nature. Thus, the aforementioned assays are usefulin determining the therapeutic utility of compounds of the invention,for example, their utility as antihypertensive agents. Other propertiesand utilities of compounds of the invention can be demonstrated usingvarious in vitro and in vivo assays well-known to those skilled in theart.

Compounds of the invention are expected to be useful for the treatmentand/or prevention of medical conditions responsive to AT₁ receptorantagonism and/or NEP inhibition. Thus it is expected that patientssuffering from a disease or disorder that is treated by antagonizing theAT₁ receptor and/or by inhibiting the NEP enzyme can be treated byadministering a therapeutically effective amount of a compound of theinvention. For example, by antagonizing the AT₁ receptor and thusinterfering with the action of angiotensin II on its receptors, thesecompounds are expected to find utility in preventing the increase inblood pressure produced by angiotensin II, a potent vasopressor. Inaddition, by inhibiting NEP, the compounds are also expected topotentiate the biological effects of endogenous peptides that aremetabolized by NEP, such as the natriuretic peptides, bombesin,bradykinins, calcitonin, endothelins, enkephalins, neurotensin,substance P and vasoactive intestinal peptide. For example, bypotentiating the effects of the natriuretic peptides, compounds of theinvention are expected to be useful to treat glaucoma. These compoundsare also expected to have other physiological actions, for example, onthe renal, central nervous, reproductive and gastrointestinal systems.

Compounds of the invention are expected to find utility in treatingand/or preventing medical conditions such as cardiovascular and renaldiseases. Cardiovascular diseases of particular interest include heartfailure such as congestive heart failure, acute heart failure, chronicheart failure, and acute and chronic decompensated heart failure. Renaldiseases of particular interest include diabetic nephropathy and chronickidney disease. One embodiment of the invention relates to a method fortreating hypertension, comprising administering to a patient atherapeutically effective amount of a compound of the invention.Typically, the therapeutically effective amount is the amount that issufficient to lower the patient's blood pressure. In one embodiment, thecompound is administered as an oral dosage form.

Another embodiment of the invention relates to a method for treatingheart failure, comprising administering to a patient a therapeuticallyeffective amount of a compound of the invention. Typically, thetherapeutically effective amount is the amount that is sufficient tolower blood pressure and/or improve renal functions. In one embodiment,the compound is administered as an intravenous dosage form. When used totreat heart failure, the compound may be administered in combinationwith other therapeutic agents such as diuretics, natriuretic peptides,and adenosine receptors antagonist.

Compounds of the invention are also expected to be useful inpreventative therapy, for example in preventing the progression ofcardiac insufficiency after myocardial infarction, preventing arterialrestenosis after angioplasty, preventing thickening of blood vesselwalls after vascular operations, preventing atherosclerosis, andpreventing diabetic angiopathy.

In addition, as NEP inhibitors, compounds of the invention are expectedto inhibit enkephalinase, which will inhibit the degradation ofendogenous enkephalins. Thus, such compounds may also find utility asanalgesics. Due to their NEP inhibition properties, compounds of theinvention are also expected to be useful as antitussive agents andantidiarrheal agents (for example, for the treatment of waterydiarrhea), as well as find utility in the treatment of menstrualdisorders, preterm labor, pre-eclampsia, endometriosis, reproductivedisorders (e.g., male and female infertility, polycystic ovariansyndrome, implantation failure), and male and female sexual dysfunction,including male erectile dysfunction and female sexual arousal disorder.More specifically, the compounds of the invention are expected to beuseful in treating female sexual dysfunction, which is often defined asa female patient's difficulty or inability to find satisfaction insexual expression. This covers a variety of diverse female sexualdisorders including, by way of illustration and not limitation,hypoactive sexual desire disorder, sexual arousal disorder, orgasmicdisorders and sexual pain disorders. When used to treat such disorders,especially female sexual dysfunction, the compounds of the invention maybe combined with one or more of the following secondary agents: PDE5inhibitors, dopamine agonists, estrogen receptor agonists and/orantagonists, androgens, and estrogens.

The amount of the compound of the invention administered per dose or thetotal amount administered per day may be predetermined or it may bedetermined on an individual patient basis by taking into considerationnumerous factors, including the nature and severity of the patient'scondition, the condition being treated, the age, weight, and generalhealth of the patient, the tolerance of the patient to the active agent,the route of administration, pharmacological considerations such as theactivity, efficacy, pharmacokinetics and toxicology profiles of thecompound and any secondary agents being administered, and the like.Treatment of a patient suffering from a disease or medical condition(such as hypertension) can begin with a predetermined dosage or a dosagedetermined by the treating physician, and will continue for a period oftime necessary to prevent, ameliorate, suppress, or alleviate thesymptoms of the disease or medical condition. Patients undergoing suchtreatment will typically be monitored on a routine basis to determinethe effectiveness of therapy. For example, in treating hypertension,blood pressure measurements may be used to determine the effectivenessof treatment. Similar indicators for other diseases and conditionsdescribed herein, are well-known and are readily available to thetreating physician. Continuous monitoring by the physician will insurethat the optimal amount of the compound of the invention will beadministered at any given time, as well as facilitating thedetermination of the duration of treatment. This is of particular valuewhen secondary agents are also being administered, as their selection,dosage, and duration of therapy may also require adjustment. In thisway, the treatment regimen and dosing schedule can be adjusted over thecourse of therapy so that the lowest amount of active agent thatexhibits the desired effectiveness is administered and, further, thatadministration is continued only so long as is necessary to successfullytreat the disease or medical condition.

Since compounds of the invention possess AT₁ receptor antagonistactivity and/or NEP enzyme inhibition activity, such compounds are alsouseful as research tools for investigating or studying biologicalsystems or samples having AT₁ receptors or a NEP enzyme, for example tostudy diseases where the AT₁ receptor or NEP enzyme plays a role. Anysuitable biological system or sample having AT₁ receptors and/or a NEPenzyme may be employed in such studies which may be conducted either invitro or in vivo. Representative biological systems or samples suitablefor such studies include, but are not limited to, cells, cellularextracts, plasma membranes, tissue samples, isolated organs, mammals(such as mice, rats, guinea pigs, rabbits, dogs, pigs, humans, and soforth), and the like, with mammals being of particular interest. In oneparticular embodiment of the invention an AT₁ receptor in a mammal isantagonized by administering an AT₁-antagonizing amount of a compound ofthe invention. In another particular embodiment, NEP enzyme activity ina mammal is inhibited by administering a NEP-inhibiting amount of acompound of the invention. Compounds of the invention can also be usedas research tools by conducting biological assays using such compounds.

When used as a research tool, a biological system or sample comprisingan AT₁ receptor and/or a NEP enzyme is typically contacted with an AT₁receptor-antagonizing or NEP enzyme-inhibiting amount of a compound ofthe invention. After the biological system or sample is exposed to thecompound, the effects of antagonizing the AT₁ receptor and/or inhibitingthe NEP enzyme are determined using conventional procedures andequipment, such as by measuring receptor binding in a binding assay ormeasuring ligand-mediated changes in a functional assay. Exposureencompasses contacting cells or tissue with the compound, administeringthe compound to a mammal, for example by i.p., i.v. or s.c.administration, and so forth. This determining step can involvemeasuring a response (a quantitative analysis) or can involve making anobservation (a qualitative analysis). Measuring a response involves, forexample, determining the effects of the compound on the biologicalsystem or sample using conventional procedures and equipment, such asradioligand binding assays and measuring ligand-mediated changes infunctional assays. The assay results can be used to determine theactivity level as well as the amount of compound necessary to achievethe desired result, i.e., an AT₁ receptor-antagonizing and/or a NEPenzyme-inhibiting amount. Typically, the determining step will involvedetermining the AT₁ receptor ligand-mediated effects and/or determiningthe effects of inhibiting the NEP enzyme.

Additionally, compounds of the invention can be used as research toolsfor evaluating other chemical compounds, and thus are also useful inscreening assays to discover, for example, new compounds having AT₁receptor-antagonizing activity and/or NEP-inhibiting activity. In thismanner, a compound of the invention is used as a standard in an assay toallow comparison of the results obtained with a test compound and withcompounds of the invention to identify those test compounds that haveabout equal or superior activity, if any. For example, K_(i) data (asdetermined, for example, by a binding assay) for a test compound or agroup of test compounds is compared to the K_(i) data for a compound ofthe invention to identify those test compounds that have the desiredproperties, e.g., test compounds having a K_(i) value about equal orsuperior to a compound of the invention, if any. This aspect of theinvention includes, as separate embodiments, both the generation ofcomparison data (using the appropriate assays) and the analysis of thetest data to identify test compounds of interest. Thus, a test compoundcan be evaluated in a biological assay, by a method comprising the stepsof: (a) conducting a biological assay with a test compound to provide afirst assay value; (b) conducting the biological assay with a compoundof the invention to provide a second assay value; wherein step (a) isconducted either before, after or concurrently with step (b); and (c)comparing the first assay value from step (a) with the second assayvalue from step (b). Exemplary biological assays include an AT₁ receptorbinding assay and a NEP enzyme inhibition assay.

Pharmaceutical Compositions and Formulations

Compounds of the invention are typically administered to a patient inthe form of a pharmaceutical composition or formulation. Suchpharmaceutical compositions may be administered to the patient by anyacceptable route of administration including, but not limited to, oral,rectal, vaginal, nasal, inhaled, topical (including transdermal),ocular, and parenteral modes of administration. Further, the compoundsof the invention may be administered, for example orally, in multipledoses per day (e.g., two, three, or four times daily), in a single dailydose or a single weekly dose. It will be understood that any form of thecompounds of the invention, (i.e., free base, free acid,pharmaceutically acceptable salt, solvate, etc.) that is suitable forthe particular mode of administration can be used in the pharmaceuticalcompositions discussed herein.

Accordingly, in one embodiment, the invention relates to apharmaceutical composition comprising a pharmaceutically acceptablecarrier and a compound of the invention. The compositions may containother therapeutic and/or formulating agents if desired. When discussingcompositions, the “compound of the invention” may also be referred toherein as the “active agent,” to distinguish it from other components ofthe formulation, such as the carrier. Thus, it is understood that theterm “active agent” includes compounds of formula I as well aspharmaceutically acceptable salts, solvates and prodrugs of thatcompound.

The pharmaceutical compositions of the invention typically contain atherapeutically effective amount of a compound of the invention. Thoseskilled in the art will recognize, however, that a pharmaceuticalcomposition may contain more than a therapeutically effective amountsuch as in bulk compositions, or less than a therapeutically effectiveamount such as in individual unit doses designed for multipleadministration to achieve a therapeutically effective amount. Typically,the composition will contain from about 0.01-95 wt % of active agent,including, from about 0.01-30 wt %, such as from about 0.01-10 wt %,with the actual amount depending upon the formulation itself, the routeof administration, the frequency of dosing, and so forth. In oneembodiment, a composition suitable for an oral dosage form, for example,may contain about 5-70 wt %, or from about 10-60 wt % of active agent.

Any conventional carrier or excipient may be used in the pharmaceuticalcompositions of the invention. The choice of a particular carrier orexcipient, or combinations of carriers or excipients, will depend on themode of administration being used to treat a particular patient or typeof medical condition or disease state. In this regard, the preparationof a suitable composition for a particular mode of administration iswell within the scope of those skilled in the pharmaceutical arts.Additionally, carriers or excipients used in such compositions arecommercially available. By way of further illustration, conventionalformulation techniques are described in Remington: The Science andPractice of Pharmacy, 20^(th) Edition, Lippincott Williams & White,Baltimore, Md. (2000); and H. C. Ansel et al., Pharmaceutical DosageForms and Drug Delivery Systems, 7^(th) Edition, Lippincott Williams &White, Baltimore, Md. (1999).

Representative examples of materials which can serve as pharmaceuticallyacceptable carriers include, but are not limited to, the following:sugars, such as lactose, glucose and sucrose; starches, such as cornstarch and potato starch; cellulose, such as microcrystalline cellulose,and its derivatives, such as sodium carboxymethyl cellulose, ethylcellulose and cellulose acetate; powdered tragacanth; malt; gelatin;talc; excipients, such as cocoa butter and suppository waxes; oils, suchas peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil,corn oil and soybean oil; glycols, such as propylene glycol; polyols,such as glycerin, sorbitol, mannitol and polyethylene glycol; esters,such as ethyl oleate and ethyl laurate; agar; buffering agents, such asmagnesium hydroxide and aluminum hydroxide; alginic acid; pyrogen-freewater; isotonic saline; Ringer's solution; ethyl alcohol; phosphatebuffer solutions; compressed propellant gases, such aschlorofluorocarbons and hydrofluorocarbons; and other non-toxiccompatible substances employed in pharmaceutical compositions.

Pharmaceutical compositions are typically prepared by thoroughly andintimately mixing or blending the active agent with a pharmaceuticallyacceptable carrier and one or more optional ingredients. The resultinguniformly blended mixture may then be shaped or loaded into tablets,capsules, pills, canisters, cartridges, dispensers and the like usingconventional procedures and equipment.

In those formulations where the compound of the invention contains athiol group, additional consideration may be given to minimize oreliminate oxidation of the thiol to form a disulfide. In solidformulations, this may be accomplished by reducing the drying time,decreasing the moisture content of the formulation, and includingmaterials such as ascorbic acid, sodium ascorbate, sodium sulfite andsodium bisulfite, as well as materials such as a mixture of lactose andmicrocrystalline cellulose. In liquid formulations, stability of thethiol may be improved by the addition of amino acids, antioxidants, or acombination of disodium edetate and ascorbic acid.

In one embodiment, the pharmaceutical compositions are suitable for oraladministration. Suitable compositions for oral administration may be inthe form of capsules, tablets, pills, lozenges, cachets, dragees,powders, granules; solutions or suspensions in an aqueous or non-aqueousliquid; oil-in-water or water-in-oil liquid emulsions; elixirs orsyrups; and the like; each containing a predetermined amount of theactive agent.

When intended for oral administration in a solid dosage form (i.e., ascapsules, tablets, pills and the like), the composition will typicallycomprise the active agent and one or more pharmaceutically acceptablecarriers, such as sodium citrate or dicalcium phosphate. Solid dosageforms may also comprise: fillers or extenders, such as starches,microcrystalline cellulose, lactose, sucrose, glucose, mannitol, and/orsilicic acid; binders, such as carboxymethylcellulose, alginates,gelatin, polyvinyl pyrrolidone, sucrose and/or acacia; humectants, suchas glycerol; disintegrating agents, such as agar-agar, calciumcarbonate, potato or tapioca starch, alginic acid, certain silicates,and/or sodium carbonate; solution retarding agents, such as paraffin;absorption accelerators, such as quaternary ammonium compounds; wettingagents, such as cetyl alcohol and/or glycerol monostearate; absorbents,such as kaolin and/or bentonite clay; lubricants, such as talc, calciumstearate, magnesium stearate, solid polyethylene glycols, sodium laurylsulfate, and/or mixtures thereof, coloring agents; and buffering agents.

Release agents, wetting agents, coating agents, sweetening, flavoringand perfuming agents, preservatives and antioxidants may also be presentin the pharmaceutical compositions. Exemplary coating agents fortablets, capsules, pills and like, include those used for entericcoatings, such as cellulose acetate phthalate, polyvinyl acetatephthalate, hydroxypropyl methylcellulose phthalate, methacrylicacid-methacrylic acid ester copolymers, cellulose acetate trimellitate,carboxymethyl ethyl cellulose, hydroxypropyl methyl cellulose acetatesuccinate, and the like. Examples of pharmaceutically acceptableantioxidants include: water-soluble antioxidants, such as ascorbic acid,cysteine hydrochloride, sodium bisulfate, sodium metabisulfate sodiumsulfite and the like; oil-soluble antioxidants, such as ascorbylpalmitate, butylated hydroxyanisole, butylated hydroxytoluene, lecithin,propyl gallate, alpha-tocopherol, and the like; and metal-chelatingagents, such as citric acid, ethylenediamine tetraacetic acid, sorbitol,tartaric acid, phosphoric acid, and the like.

Compositions may also be formulated to provide slow or controlledrelease of the active agent using, by way of example, hydroxypropylmethyl cellulose in varying proportions or other polymer matrices,liposomes and/or microspheres. In addition, the pharmaceuticalcompositions of the invention may contain opacifying agents and may beformulated so that they release the active agent only, orpreferentially, in a certain portion of the gastrointestinal tract,optionally, in a delayed manner. Examples of embedding compositionswhich can be used include polymeric substances and waxes. The activeagent can also be in micro-encapsulated form, if appropriate, with oneor more of the above-described excipients.

Suitable liquid dosage forms for oral administration include, by way ofillustration, pharmaceutically acceptable emulsions, microemulsions,solutions, suspensions, syrups and elixirs. Liquid dosage formstypically comprise the active agent and an inert diluent, such as, forexample, water or other solvents, solubilizing agents and emulsifiers,such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethylacetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butyleneglycol, oils (e.g., cottonseed, groundnut, corn, germ, olive, castor andsesame oils), glycerol, tetrahydrofuryl alcohol, polyethylene glycolsand fatty acid esters of sorbitan, and mixtures thereof. Suspensions maycontain suspending agents such as, for example, ethoxylated isostearylalcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystallinecellulose, aluminium metahydroxide, bentonite, agar-agar and tragacanth,and mixtures thereof.

When intended for oral administration, the pharmaceutical compositionsof the invention may be packaged in a unit dosage form. The term “unitdosage form” refers to a physically discrete unit suitable for dosing apatient, i.e., each unit containing a predetermined quantity of theactive agent calculated to produce the desired therapeutic effect eitheralone or in combination with one or more additional units. For example,such unit dosage forms may be capsules, tablets, pills, and the like.

In another embodiment, the compositions of the invention are suitablefor inhaled administration, and will typically be in the form of anaerosol or a powder. Such compositions are generally administered usingwell-known delivery devices, such as a nebulizer, dry powder, ormetered-dose inhaler. Nebulizer devices produce a stream of highvelocity air that causes the composition to spray as a mist that iscarried into a patient's respiratory tract. An exemplary nebulizerformulation comprises the active agent dissolved in a carrier to form asolution, or micronized and combined with a carrier to form a suspensionof micronized particles of respirable size. Dry powder inhalersadminister the active agent as a free-flowing powder that is dispersedin a patient's air-stream during inspiration. An exemplary dry powderformulation comprises the active agent dry-blended with an excipientsuch as lactose, starch, mannitol, dextrose, polylactic acid,polylactide-co-glycolide, and combinations thereof. Metered-doseinhalers discharge a measured amount of the active agent usingcompressed propellant gas. An exemplary metered-dose formulationcomprises a solution or suspension of the active agent in a liquefiedpropellant, such as a chlorofluorocarbon or hydrofluoroalkane. Optionalcomponents of such formulations include co-solvents, such as ethanol orpentane, and surfactants, such as sorbitan trioleate, oleic acid,lecithin, glycerin, and sodium lauryl sulfate. Such compositions aretypically prepared by adding chilled or pressurized hydrofluoroalkane toa suitable container containing the active agent, ethanol (if present)and the surfactant (if present). To prepare a suspension, the activeagent is micronized and then combined with the propellant.Alternatively, a suspension formulation can be prepared by spray dryinga coating of surfactant on micronized particles of the active agent. Theformulation is then loaded into an aerosol canister, which forms aportion of the inhaler.

Compounds of the invention can also be administered parenterally (e.g.,by subcutaneous, intravenous, intramuscular, or intraperitonealinjection). For such administration, the active agent is provided in asterile solution, suspension, or emulsion. Exemplary solvents forpreparing such formulations include water, saline, low molecular weightalcohols such as propylene glycol, polyethylene glycol, oils, gelatin,fatty acid esters such as ethyl oleate, and the like. Parenteralformulations may also contain one or more anti-oxidants, solubilizers,stabilizers, preservatives, wetting agents, emulsifiers, and dispersingagents. Surfactants, additional stabilizing agents or pH-adjustingagents (acids, bases or buffers) and anti-oxidants are particularlyuseful to provide stability to the formulation, for example, to minimizeor avoid hydrolysis of ester and amide linkages, or dimerization ofthiols that may be present in the compound. These formulations may berendered sterile by use of a sterile injectable medium, a sterilizingagent, filtration, irradiation, or heat. In one particular embodiment,the parenteral formulation comprises an aqueous cyclodextrin solution asthe pharmaceutically acceptable carrier. Suitable cyclodextrins includecyclic molecules containing six or more α-D-glucopyranose units linkedat the 1,4 positions by a linkages as in amylase, β-cyclodextrin orcycloheptaamylose. Exemplary cyclodextrins include cyclodextrinderivatives such as hydroxypropyl and sulfobutyl ether cyclodextrinssuch as hydroxypropyl-β-cyclodextrin and sulfobutyl etherβ-cyclodextrin. Exemplary buffers for such formulations includecarboxylic acid-based buffers such as citrate, lactate and maleatebuffer solutions.

Compounds of the invention can also be administered transdermally usingknown transdermal delivery systems and excipients. For example, thecompound can be admixed with permeation enhancers, such as propyleneglycol, polyethylene glycol monolaurate, azacycloalkan-2-ones and thelike, and incorporated into a patch or similar delivery system.Additional excipients including gelling agents, emulsifiers and buffers,may be used in such transdermal compositions if desired.

If desired, the compounds of the invention may be administered incombination with one or more other therapeutic agents. Thus, in oneembodiment, pharmaceutical compositions of the invention contain otherdrugs that are co-administered with a compound of the invention. Forexample, the composition may further comprise one or more drugs (alsoreferred to as “secondary agents(s)”) selected from the group ofdiuretics, β₁ adrenergic receptor blockers, calcium channel blockers,angiotensin-converting enzyme inhibitors, AT₁ receptor antagonists,neprilysin inhibitors, non-steroidal anti-inflammatory agents,prostaglandins, anti-lipid agents, anti-diabetic agents, anti-thromboticagents, renin inhibitors, endothelin receptor antagonists, endothelinconverting enzyme inhibitors, aldosterone antagonists,angiotensin-converting enzyme/neprilysin inhibitors, and combinationsthereof. Such therapeutic agents are well known in the art, and specificexamples are described herein. By combining a compound of the inventionwith a secondary agent, triple therapy can be achieved, i.e., AT₁receptor antagonist activity, NEP inhibition activity and activityassociated with the secondary agent (e.g., β₁ adrenergic receptorblocker), using only two active components. Since compositionscontaining two active components are typically easier to formulate thancompositions containing three active components, such two-componentcompositions provide a significant advantage over compositionscontaining three active components. Accordingly, in yet another aspectof the invention, a pharmaceutical composition comprises a compound ofthe invention, a second active agent, and a pharmaceutically acceptablecarrier. Third, fourth, etc., active agents may also be included in thecomposition. In combination therapy, the amount of compound of theinvention that is administered, as well as the amount of secondaryagents, may be less than the amount typically administered inmonotherapy.

Compounds of the invention may be physically mixed with the secondactive agent to form a composition containing both agents; or each agentmay be present in separate and distinct compositions which areadministered to the patient simultaneously or at separate times. Forexample, a compound of the invention can be combined with a secondactive agent using conventional procedures and equipment to form acombination of active agents comprising a compound of the invention anda second active agent. Additionally, the active agents may be combinedwith a pharmaceutically acceptable carrier to form a pharmaceuticalcomposition comprising a compound of the invention, a second activeagent and a pharmaceutically acceptable carrier. In this embodiment, thecomponents of the composition are typically mixed or blended to create aphysical mixture. The physical mixture is then administered in atherapeutically effective amount using any of the routes describedherein.

Alternatively, the active agents may remain separate and distinct beforeadministration to the patient. In this embodiment, the agents are notphysically mixed together before administration but are administeredsimultaneously or at separate times as separate compositions. Suchcompositions can be packaged separately or may be packaged together in akit. When administered at separate times, the secondary agent willtypically be administered less than 24 hours after administration of thecompound of the invention, ranging anywhere from concurrent withadministration of the compound of the invention to about 24 hourspost-dose. This is also referred to as sequential administration. Thus,a compound of the invention can be orally administered simultaneously orsequentially with another active agent using two tablets, with onetablet for each active agent, where sequential may mean beingadministered immediately after administration of the compound of theinvention or at some predetermined time later (e.g., one hour later orthree hours later). Alternatively, the combination may be administeredby different routes of administration, i.e., one orally and the other byinhalation.

In one embodiment, the kit comprises a first dosage form comprising acompound of the invention and at least one additional dosage formcomprising one or more of the secondary agents set forth herein, inquantities sufficient to carry out the methods of the invention. Thefirst dosage form and the second (or third, etc,) dosage form togethercomprise a therapeutically effective amount of active agents for thetreatment or prevention of a disease or medical condition in a patient.

Secondary agent(s), when included, are present in a therapeuticallyeffective amount such that they produce a therapeutically beneficialeffect when co-administered with a compound of the invention. Thesecondary agent can be in the form of a pharmaceutically acceptablesalt, solvate, optically pure stereoisomer, and so forth. The secondaryagent may also be in the form of a prodrug, for example, a compoundhaving a carboxylic acid group that has been esterified. Thus, secondaryagents listed herein are intended to include all such forms, and arecommercially available or can be prepared using conventional proceduresand reagents.

In one embodiment, a compound of the invention is administered incombination with a diuretic. Representative diuretics include, but arenot limited to: carbonic anhydrase inhibitors such as acetazolamide anddichlorphenamide; loop diuretics, which include sulfonamide derivativessuch as acetazolamide, ambuside, azosemide, bumetanide, butazolamide,chloraminophenamide, clofenamide, clopamide, clorexolone, disulfamide,ethoxolamide, furosemide, mefruside, methazolamide, piretanide,torsemide, tripamide, and xipamide, as well as non-sulfonamide diureticssuch as ethacrynic acid and other phenoxyacetic acid compounds such astienilic acid, indacrinone and quincarbate; osmotic diuretics such asmannitol; potassium-sparing diuretics, which include aldosteroneantagonists such as spironolactone, and Na⁺ channel inhibitors such asamiloride and triamterene; thiazide and thiazide-like diuretics such asalthiazide, bendroflumethiazide, benzylhydrochlorothiazide,benzthiazide, buthiazide, chlorthalidone, chlorothiazide,cyclopenthiazide, cyclothiazide, epithiazide, ethiazide, fenquizone,flumethiazide, hydrochlorothiazide, hydroflumethiazide, indapamide,methylclothiazide, meticrane, metolazone, paraflutizide, polythiazide,quinethazone, teclothiazide, and trichloromethiazide; and combinationsthereof. In a particular embodiment, the diuretic is selected fromamiloride, bumetanide, chlorothiazide, chlorthalidone, dichlorphenamide,ethacrynic acid, furosemide, hydrochlorothiazide, hydroflumethiazide,indapamide, methylclothiazide, metolazone, torsemide, triamterene, andcombinations thereof. The diuretic will be administered in an amountsufficient to provide from about 5-50 mg per day, more typically 6-25 mgper day, with common dosages being 6.25 mg, 12.5 mg or 25 mg per day.

Compounds of the invention may also be administered in combination witha β₁ adrenergic receptor blocker. Representative β₁ adrenergic receptorblockers include, but are not limited to, acebutolol, alprenolol,amosulalol, arotinolol, atenolol, befunolol, betaxolol, bevantolol,bisoprolol, bopindolol, bucindolol, bucumolol, bufetolol, bufuralol,bunitrolol, bupranolol, bubridine, butofilolol, carazolol, carteolol,carvedilol, celiprolol, cetamolol, cloranolol, dilevalol, epanolol,esmolol, indenolol, labetolol, levobunolol, mepindolol, metipranolol,metoprolol such as metoprolol succinate and metoprolol tartrate,moprolol, nadolol, nadoxolol, nebivalol, nipradilol, oxprenolol,penbutolol, perbutolol, pindolol, practolol, pronethalol, propranolol,sotalol, sufinalol, talindol, tertatolol, tilisolol, timolol,toliprolol, xibenolol, and combinations thereof. In one particularembodiment, the β₁ adrenergic receptor blocker is selected fromatenolol, bisoprolol, metoprolol, propranolol, sotalol, and combinationsthereof.

In one embodiment, a compound of the invention is administered incombination with a calcium channel blocker. Representative calciumchannel blockers include, but are not limited to, amlodipine, anipamil,aranipine, barnidipine, bencyclane, benidipine, bepridil, clentiazem,cilnidipine, cinnarizine, diltiazem, efonidipine, elgodipine, etafenone,felodipine, fendiline, flunarizine, gallopamil, isradipine, lacidipine,lercanidipine, lidoflazine, lomerizine, manidipine, mibefradil,nicardipine, nifedipine, niguldipine, niludipine, nilvadipine,nimodipine, nisoldipine, nitrendipine, nivaldipine, perhexyline,prenylamine, ryosidine, semotiadil, terodiline, tiapamil, verapamil, andcombinations thereof. In a particular embodiment, the calcium channelblocker is selected from amlodipine, bepridil, diltiazem, felodipine,isradipine, lacidipine, nicardipine, nifedipine, niguldipine,niludipine, nimodipine, nisoldipine, ryosidine, verapamil, andcombinations thereof.

Compounds of the invention can also be administered in combination withan angiotensin-converting enzyme (ACE) inhibitor. Representative ACEinhibitors include, but are not limited to, accupril, alacepril,benazepril, benazeprilat, captopril, ceranapril, cilazapril, delapril,enalapril, enalaprilat, fosinopril, fosinoprilat, imidapril, lisinopril,moexipril, monopril, moveltopril, pentopril, perindopril, quinapril,quinaprilat, ramipril, ramiprilat, saralasin acetate, spirapril,temocapril, trandolapril, zofenopril, and combinations thereof. In aparticular embodiment, the ACE inhibitor is selected from: benazepril,enalapril, lisinopril, ramipril, and combinations thereof.

In one embodiment, a compound of the invention is administered incombination with an AT₁ receptor antagonist, also known as angiotensinII type 1 receptor blockers (ARBs). Representative ARBs include, but arenot limited to, abitesartan, benzyllosartan, candesartan, candesartancilexetil, elisartan, embusartan, enoltasosartan, eprosartan, fonsartan,forasartan, glycyllosartan, irbesartan, isoteoline, losartan, medoximil,milfasartan, olmesartan, opomisartan, pratosartan, ripisartan,saprisartan, saralasin, sarmesin, tasosartan, telmisartan, valsartan,zolasartan, and combinations thereof. In a particular embodiment, theARB is selected from candesartan, eprosartan, irbesartan, losartan,olmesartan, saprisartan, tasosartan, telmisartan, valsartan, andcombinations thereof. Exemplary salts include eprosartan mesylate,losartan potassium salt, and olmesartan medoxomil. Typically, the ARBwill be administered in an amount sufficient to provide from about 4-600mg per dose, with exemplary daily dosages ranging from 20-320 mg perday.

In another embodiment, a compound of the invention is administered incombination with a neprilysin (NEP) inhibitor. Representative NEPinhibitors include, but are not limited to: candoxatril; candoxatrilat;dexecadotril ((+)-N-[2(R)-(acetylthiomethyl)-3-phenylpropionyl]glycinebenzyl ester); CGS-24128(3-[3-(biphenyl-4-yl)-2-(phosphonomethylamino)propionamido]propionicacid); CGS-24592((S)-3-[3-(biphenyl-4-yl)-2-(phosphonomethylamino)propionamido]propionicacid); CGS-25155 (N-[9(R)-(acetylthiomethyl)-10-oxo-1-azacyclodecan-2(S)-ylcarbonyl]-4(R)-hydroxy-L-proline benzyl ester);3-(1-carbamoylcyclohexyl)propionic acid derivatives described in WO2006/027680 to Hepworth et al. (Pfizer Inc.); JMV-390-1(2(R)-benzyl-3-(N-hydroxycarbamoyl)propionyl-L-isoleucyl-L-leucine);ecadotril; phosphoramidon; retrothiorphan; RU-42827(2-(mercaptomethyl)-N-(4-pyridinyl)benzenepropionamide); RU-44004(N-(4-morpholinyl)-3-phenyl-2-(sulfanylmethyl)propionamide); SCH-32615((S)—N—[N-(1-carboxy-2-phenylethyl)-L-phenylalanyl]-β-alanine) and itsprodrug SCH-34826((S)—N—[N-[1-[[(2,2-dimethyl-1,3-dioxolan-4-yl)methoxy]carbonyl]-2-phenylethyl]-L-phenylalanyl]-β-alanine);sialorphin; SCH-42495(N-[2(S)-(acetylsulfanylmethyl)-3-(2-methylphenyl)propionyl]-L-methionineethyl ester); spinorphin; SQ-28132(N-[2-(mercaptomethyl)-1-oxo-3-phenylpropyl]leucine); SQ-28603(N-[2-(mercaptomethyl)-1-oxo-3-phenylpropyl]-β-alanine); SQ-29072(7-[[2-(mercaptomethyl)-1-oxo-3-phenylpropyl]amino]heptanoic acid);thiorphan and its prodrug racecadotril; UK-69578(cis-4-[[[1-[2-carboxy-3-(2-methoxyethoxy)propyl]cyclopentyl]carbonyl]amino]cyclohexanecarboxylicacid); UK-447,841(2-{1-[3-(4-chlorophenyl)propylcarbamoyl]-cyclopentylmethyl}-4-methoxybutyricacid); UK-505,749((R)-2-methyl-3-{1-[3-(2-methylbenzothiazol-6-yl)propylcarbamoyl]cyclopentyl}propionicacid); 5-biphenyl-4-yl-4-(3-carboxypropionylamino)-2-methylpentanoicacid and 5-biphenyl-4-yl-4-(3-carboxypropionylamino)-2-methylpentanoicacid ethyl ester (WO 2007/056546); and combinations thereof. In aparticular embodiment, the NEP inhibitor is selected from candoxatril,candoxatrilat, CGS-24128, phosphoramidon, SCH-32615, SCH-34826,SQ-28603, thiorphan, and combinations thereof. The NEP inhibitor will beadministered in an amount sufficient to provide from about 20-800 mg perday, with typical daily dosages ranging from 50-700 mg per day, morecommonly 100-600 or 100-300 mg per day.

In yet another embodiment, a compound of the invention is administeredin combination with a non-steroidal anti-inflammatory agent (NSAID).Representative NSAIDs include, but are not limited to: acemetacin,acetyl salicylic acid, alclofenac, alminoprofen, amfenac, amiprilose,amoxiprin, anirolac, apazone, azapropazone, benorilate, benoxaprofen,bezpiperylon, broperamole, bucloxic acid, carprofen, clidanac,diclofenac, diflunisal, diftalone, enolicam, etodolac, etoricoxib,fenbufen, fenclofenac, fenclozic acid, fenoprofen, fentiazac, feprazone,flufenamic acid, flufenisal, fluprofen, flurbiprofen, furofenac,ibufenac, ibuprofen, indomethacin, indoprofen, isoxepac, isoxicam,ketoprofen, ketorolac, lofemizole, lornoxicam, meclofenamate,meclofenamic acid, mefenamic acid, meloxicam, mesalamine, miroprofen,mofebutazone, nabumetone, naproxen, niflumic acid, oxaprozin, oxpinac,oxyphenbutazone, phenylbutazone, piroxicam, pirprofen, pranoprofen,salsalate, sudoxicam, sulfasalazine, sulindac, suprofen, tenoxicam,tiopinac, tiaprofenic acid, tioxaprofen, tolfenamic acid, tolmetin,triflumidate, zidometacin, zomepirac, and combinations thereof. In aparticular embodiment, the NSAID is selected from etodolac,flurbiprofen, ibuprofen, indomethacin, ketoprofen, ketorolac, meloxicam,naproxen, oxaprozin, piroxicam, and combinations thereof.

In yet another embodiment, a compound of the invention is administeredin combination with an anti-lipid agent. Representative anti-lipidagents include, but are not limited to, statins such as atorvastatin,fluvastatin, lovastatin, pravastatin, rosuvastatin and simvastatin;cholesteryl ester transfer proteins (CETPs); and combinations thereof.

In yet another embodiment, a compound of the invention is administeredin combination with an anti-diabetic agent. Representative anti-diabeticagents include injectable drugs as well as orally effective drugs, andcombinations thereof. Examples of injectable drugs include, but are notlimited to, insulin and insulin derivatives. Examples of orallyeffective drugs include, but are not limited to: biguanides such asmetformin; glucagon antagonists; α-glucosidase inhibitors such asacarbose and miglitol; meglitinides such as repaglinide;oxadiazolidinediones; sulfonylureas such as chlorpropamide, glimepiride,glipizide, glyburide, and tolazamide; thiazolidinediones such aspioglitazone and rosiglitazone; and combinations thereof.

In one embodiment, a compound of the invention is administered incombination with an anti-thrombotic agent. Representativeanti-thrombotic agents include, but are not limited to, aspirin,anti-platelet agents, heparin, and combinations thereof. Compounds ofthe invention may also be administered in combination with a renininhibitor, examples of which include, but are not limited to, aliskiren,enalkiren, remikiren, and combinations thereof. In another embodiment, acompound of the invention is administered in combination with anendothelin receptor antagonist, representative examples of whichinclude, but are not limited to, bosentan, darusentan, tezosentan, andcombinations thereof. Compounds of the invention may also beadministered in combination with an endothelin converting enzymeinhibitor, examples of which include, but are not limited to,phosphoramidon, CGS 26303, and combinations thereof. In yet anotherembodiment, a compound of the invention is administered in combinationwith an aldosterone antagonist. Representative aldosterone antagonistsinclude, but are not limited to, eplerenone, spironolactone, andcombinations thereof.

Combined therapeutic agents may also be helpful in further combinationtherapy with compounds of the invention. For example, a combination ofthe ACE inhibitor enalapril (in the maleate salt form) and the diuretichydrochlorothiazide, which is sold under the mark Vaseretic®, or acombination of the calcium channel blocker amlodipine (in the besylatesalt form) and the ARB olmesartan (in the medoxomil prodrug form), or acombination of a calcium channel blocker and a statin, all may also beused with the compounds of the invention. Dual-acting agents may also behelpful in combination therapy with compounds of the invention. Forexample, angiotensin-converting enzyme/neprilysin (ACE/NEP) inhibitorssuch as: AVE-0848((4S,7S,12bR)-7-[3-methyl-2(S)-sulfanylbutyramido]-6-oxo-1,2,3,4,6,7,8,12b-octahydropyrido[2,1-a][2]benzazepine-4-carboxylicacid); AVE-7688 (ilepatril) and its parent compound; BMS-182657(2-[2-oxo-3(S)-[3-phenyl-2(S)-sulfanylpropionamido]-2,3,4,5-tetrahydro-1H-1-benzazepin-1-yl]aceticacid); CGS-26303([N-[2-(biphenyl-4-yl)-1(S)-(1H-tetrazol-5-yl)ethyl]amino]methylphosphonicacid); CGS-35601(N-[1-[4-methyl-2(S)-sulfanylpentanamido]cyclopentylcarbonyl]-L-tryptophan);fasidotril; fasidotrilate; enalaprilat; ER-32935((3R,6S,9aR)-6-[3(S)-methyl-2(S)-sulfanylpentanamido]-5-oxoperhydrothiazolo[3,2-a]azepine-3-carboxylicacid); gempatrilat; MDL-101264((4S,7S,12bR)-7-[2(S)-(2-morpholinoacetylthio)-3-phenylpropionamido]-6-oxo-1,2,3,4,6,7,8,12b-octahydropyrido[2,1-a][2]benzazepine-4-carboxylicacid); MDL-101287 ([4S-[4α,7α(R*),12bβ]]-7-[2-(carboxymethyl)-3-phenylpropionamido]-6-oxo-1,2,3,4,6,7,8,12b-octahydropyrido[2,1-a][2]benzazepine-4-carboxylicacid); omapatrilat; RB-105 (N-[2 (S)-(mercaptomethyl)-3(R)-phenylbutyl]-L-alanine); sampatrilat; SA-898((2R,4R)—N-[2-(2-hydroxyphenyl)-3-(3-mercaptopropionyl)thiazolidin-4-ylcarbonyl]-L-phenylalanine);Sch-50690(N-[1(S)-carboxy-2-[N2-(methanesulfonyl)-L-lysylamino]ethyl]-L-valyl-L-tyrosine);and combinations thereof, may also be included. In one particularembodiment, the ACE/NEP inhibitor is selected from: AVE-7688,enalaprilat, fasidotril, fasidotrilate, omapatrilat, sampatrilat, andcombinations thereof.

Other therapeutic agents such as α₂-adrenergic receptor agonists andvasopressin receptor antagonists may also be helpful in combinationtherapy. Exemplary α₂-adrenergic receptor agonists include clonidine,dexmedetomidine, and guanfacine. Exemplary vasopressin receptorantagonists include tolvaptan.

The following formulations illustrate representative pharmaceuticalcompositions of the invention.

Exemplary Hard Gelatin Capsules for Oral Administration

A compound of the invention (50 g), spray-dried lactose (440 g) andmagnesium stearate (10 g) are thoroughly blended. The resultingcomposition is then loaded into hard gelatin capsules (500 mg ofcomposition per capsule). Alternately, a compound of the invention (20mg) is thoroughly blended with starch (89 mg), microcrystallinecellulose (89 mg) and magnesium stearate (2 mg). The mixture is thenpassed through a No. 45 mesh U.S. sieve and loaded into a hard gelatincapsule (200 mg of composition per capsule).

Exemplary Gelatin Capsule Formulation for Oral Administration

A compound of the invention (100 mg) is thoroughly blended withpolyoxyethylene sorbitan monooleate (50 mg) and starch powder (250 mg).The mixture is then loaded into a gelatin capsule (300 mg of compositionper capsule). Alternately, a compound of the invention (40 mg) isthoroughly blended with microcrystalline cellulose (Avicel PH 103; 260mg) and magnesium stearate (0.8 mg). The mixture is then loaded into agelatin capsule (Size #1, White, Opaque) (300 mg of composition percapsule).

Exemplary Tablet Formulation for Oral Administration

A compound of the invention (10 mg), starch (45 mg) and microcrystallinecellulose (35 mg) are passed through a No. 20 mesh U.S. sieve and mixedthoroughly. The granules so produced are dried at 50-60° C. and passedthrough a No. 16 mesh U.S. sieve. A solution of polyvinylpyrrolidone (4mg as a 10% solution in sterile water) is mixed with sodiumcarboxymethyl starch (4.5 mg), magnesium stearate (0.5 mg), and talc (1mg), and this mixture is then passed through a No. 16 mesh U.S. sieve.The sodium carboxymethyl starch, magnesium stearate and talc are thenadded to the granules. After mixing, the mixture is compressed on atablet machine to afford a tablet weighing 100 mg.

Alternately, a compound of the invention (250 mg) is thoroughly blendedwith microcrystalline cellulose (400 mg), silicon dioxide fumed (10 mg),and stearic acid (5 mg). The mixture is then compressed to form tablets(665 mg of composition per tablet).

Alternately, a compound of the invention (400 mg) is thoroughly blendedwith cornstarch (50 mg), croscarmellose sodium (25 mg), lactose (120mg), and magnesium stearate (5 mg). The mixture is then compressed toform a single-scored tablet (600 mg of composition per tablet).

Alternately, a compound of the invention (100 mg) is thoroughly blendedwith cornstarch (100 mg) with an aqueous solution of gelatin (20 mg).The mixture is dried and ground to a fine powder Microcrystallinecellulose (50 mg) and magnesium stearate (5 mg) are the admixed with thegelatin formulation, granulated and the resulting mixture compressed toform tablets (100 mg of active per tablet).

Exemplary Suspension Formulation for Oral Administration

The following ingredients are mixed to form a suspension containing 100mg of active agent per 10 mL of suspension:

Ingredients Amount Compound of the invention 1.0 g Fumaric acid 0.5 gSodium chloride 2.0 g Methyl paraben 0.15 g Propyl paraben 0.05 gGranulated sugar 25.5 g Sorbitol (70% solution) 12.85 g Veegum ® K(magnesium aluminum silicate) 1.0 g Flavoring 0.035 mL Colorings 0.5 mgDistilled water q.s. to 100 mL

Exemplary Liquid Formulation for Oral Administration

A suitable liquid formulation is one with a carboxylic acid-based buffersuch as citrate, lactate and maleate buffer solutions. For example, acompound of the invention (which may be pre-mixed with DMSO) is blendedwith a 100 mM ammonium citrate buffer and the pH adjusted to pH 5, orwith is blended with a 100 mM citric acid solution and the pH adjustedto pH 2. Such solutions may also include a solubilizing excipient suchas a cyclodextrin, for example the solution may include 10 wt %hydroxypropyl-β-cyclodextrin.

Exemplary Injectable Formulation for Administration by Injection

A compound of the invention (0.2 g) is blended with 0.4 M sodium acetatebuffer solution (2.0 mL). The pH of the resulting solution is adjustedto pH 4 using 0.5 N aqueous hydrochloric acid or 0.5 N aqueous sodiumhydroxide, as necessary, and then sufficient water for injection isadded to provide a total volume of 20 mL. The mixture is then filteredthrough a sterile filter (0.22 micron) to provide a sterile solutionsuitable for administration by injection.

Exemplary Compositions for Administration by Inhalation

A compound of the invention (0.2 mg) is micronized and then blended withlactose (25 mg). This blended mixture is then loaded into a gelatininhalation cartridge. The contents of the cartridge are administeredusing a dry powder inhaler, for example.

Alternately, a micronized compound of the invention (10 g) is dispersedin a solution prepared by dissolving lecithin (0.2 g) in demineralizedwater (200 mL). The resulting suspension is spray dried and thenmicronized to form a micronized composition comprising particles havinga mean diameter less than about 1.5 μm. The micronized composition isthen loaded into metered-dose inhaler cartridges containing pressurized1,1,1,2-tetrafluoroethane in an amount sufficient to provide about 10 μgto about 500 μg of the compound of the invention per dose whenadministered by the inhaler.

Alternately, a compound of the invention (25 mg) is dissolved in citratebuffered (pH 5) isotonic saline (125 mL). The mixture is stirred andsonicated until the compound is dissolved. The pH of the solution ischecked and adjusted, if necessary, to pH 5 by slowly adding aqueous 1Nsodium hydroxide. The solution is administered using a nebulizer devicethat provides about 10 μg to about 500 μg of the compound of theinvention per dose.

EXAMPLES

The following Preparations and Examples are provided to illustratespecific embodiments of the invention. These specific embodiments,however, are not intended to limit the scope of the invention in any wayunless specifically indicated.

The following abbreviations have the following meanings unless otherwiseindicated and any other abbreviations used herein and not defined havetheir standard meaning:

-   -   ACE angiotensin converting enzyme    -   APP aminopeptidase P    -   AT₁ angiotensin II type 1 (receptor)    -   AT₂ angiotensin II type 2 (receptor)    -   BCA bicinchoninic acid    -   BSA bovine serum albumin    -   DCM dichloromethane    -   DMF N,N-dimethylformamide    -   DMSO dimethyl sulfoxide    -   Dnp 2,4-dinitrophenyl    -   DOCA deoxycorticosterone acetate    -   EDC N-(3-dimethylaminopropyl)-N′-ethylcarbodiimide    -   EDTA ethylenediaminetetraacetic acid    -   EGTA ethylene glycol bis(β-aminoethyl        ether)-N,N,N′N′-tetraacetic acid    -   EtOAc ethyl acetate    -   EtOH ethanol    -   HOBt 1-hydroxybenzotriazole    -   Mca (7-methoxycoumarin-4-yl)acyl    -   MeCN acetonitrile    -   MeOH methanol    -   NBS N-bromosuccinimide    -   NEP neprilysin (EC 3.4.24.11)    -   PBS phosphate buffered saline    -   SHR spontaneously hypertensive rat    -   TFA trifluoroacetic acid    -   THF tetrahydrofuran    -   Tris tris(hydroxymethyl)aminomethane    -   Tween-20 polyethylene glycol sorbitan monolaurate

Unless noted otherwise, all materials, such as reagents, startingmaterials and solvents, were purchased from commercial suppliers (suchas Sigma-Aldrich, Fluka Riedel-de Haen, and the like) and were usedwithout further purification.

Reactions were run under nitrogen atmosphere, unless noted otherwise.The progress of reactions were monitored by thin layer chromatography(TLC), analytical high performance liquid chromatography (anal. HPLC),and mass spectrometry, the details of which are given in specificexamples. Solvents used in analytical HPLC were as follows: solvent Awas 98% water/2% MeCN/1.0 mL/L TFA; solvent B was 90% MeCN/10% water/1.0mL/L TFA.

Reactions were worked up as described specifically in each preparationor example; commonly reaction mixtures were purified by extraction andother purification methods such as temperature-, and solvent-dependentcrystallization, and precipitation. In addition, reaction mixtures wereroutinely purified by preparative HPLC, typically using Microsorb C18and Microsorb BDS column packings and conventional eluents.Characterization of reaction products was routinely carried out by massand ¹H-NMR spectrometry. For NMR measurement, samples were dissolved indeuterated solvent (CD₃OD, CDCl₃, or DMSO-d₆), and ¹H-NMR spectra wereacquired with a Varian Gemini 2000 instrument (400 MHz) under standardobservation conditions. Mass spectrometric identification of compoundswas typically conducted using an electrospray ionization method (ESMS)with an Applied Biosystems (Foster City, Calif.) model API 150 EXinstrument or an Agilent (Palo Alto, Calif.) model 1200 LC/MSDinstrument.

Preparation 1 7-Methyl-2-propyl-3H-benzoimidazole-5-carboxylic acid((R)-1-benzyl-2-benzyloxycarbamoylethyl)amide

To a solution of 7-methyl-2-propyl-3H-benzoimidazole-5-carboxylic acid(164 mg, 752 μmol) and (R)-3-amino-N-benzyloxy-4-phenylbutyramide (TFAsalt: 300 mg, 754 μmol) in DMF (10 mL) containing triethylamine (210μL), was added HOBt (151 μg, 755 μmol) and EDC (151 mg, 788 μmol). Themixture was stirred at room temperature overnight and concentrated invacuo, yielding a pale brown residue. The residue was dissolved in DCM(100 mL) and washed sequentially with 1M H₃PO₄, a saturated NaHCO₃solution, and saturated aqueous NaCl. The organic layer was collected,dried over MgSO₄, and concentrated to afford the title compound as apale yellow oil (150 mg; 41% yield), which was used without furthertreatment. ESMS [M+H]⁺ calcd for C₂₉H₃₂N₄O₃, 485.26; found 485.5.

Preparation 24-[6-((R)-1-Benzyl-2-benzyloxycarbamoylethylcarbamoyl)-4-methyl-2-propylbenzoimidazol-1-ylmethyl]benzoicAcid Methyl Ester (2a) and4-[5-((R)-1-Benzyl-2-benzyloxycarbamoylethylcarbamoyl-7-methyl-2-propylbenzoimidazol-1-ylmethyl]benzoicAcid Methyl Ester (2b)

To a cold solution of 7-methyl-2-propyl-3H-benzoimidazole-5-carboxylicacid ((R)-1-benzyl-2-benzyloxycarbamoylethyl)amide (150 mg, 310 μmol) inDMF (10 mL) in an ice bath, was added NaH (60% dispersion in mineraloil; 56 mg) under nitrogen. After stirring the mixture for 20 minutes,4-bromomethylbenzoic acid methyl ester (71 mg, 310 μmol) was added. Thefinal mixture was stirred at room temperature for 2 hours, then at 80°C. for 12 hours. The mixture was cooled, and concentrated in vacuo. Theresulting residue was washed with hexane (10 mL), dissolved in DCM (50mL), and sequentially washed with 1M H₃PO₄, a saturated NaHCO₃ solution,and saturated aqueous NaCl. The organic layer was collected and driedover MgSO₄, and concentrated in vacuo, to afford a pale yellow oil. Thecrude oil contained a mixture of two alkylation products, compound 2a (amajor product) and compound 2b (a minor desired product). ESMS [M+H]⁺calcd for C₃₈H₄₀N₄O₅, 632.30; found 633.4.

Preparation 34-[6-((R)-1-Benzyl-2-benzyloxycarbamoylethylcarbamoyl)-4-methyl-2-propylbenzoimidazol-1-ylmethyl]benzoicAcid (3a) and4-[5-((R)-1-Benzyl-2-benzyloxycarbamoylethylcarbamoyl)-7-methyl-2-propylbenzoimidazol-1-ylmethyl]benzoicAcid (3b)

A mixture of4-[6-((R)-1-benzyl-2-benzyloxycarbamoylethylcarbamoyl)-4-methyl-2-propylbenzoimidazol-1-ylmethyl]benzoicacid methyl ester and4-[5-((R)-1-benzyl-2-benzyloxycarbamoylethylcarbamoyl)-7-methyl-2-propylbenzoimidazol-1-ylmethyl]benzoicacid methyl ester was dissolved in a mixture of MeOH (20 mL) and THF (5mL), to which an aqueous NaOH solution (33 mg, 825 μmol; 1 mL) wasadded. The mixture was stirred at room temperature for 24 hours, andconcentrated in vacuo to yield a pale brown residue. The residue wassuspended in water, followed by the addition of 1M H₃PO₄ until the pH ofthe solution reached 3. The precipitated solid was collected, dissolvedin MeOH and evaporated to dryness to yield the title compounds as a paleyellow oil, which was used without further purification. ESMS [M+H]⁺calcd for C₃₇H₃₈N₄O₅, 619.29; found 619.0.

Example 14-[6-((R)-1-Benzyl-2-hydroxycarbamoylethylcarbamoyl)-4-methyl-2-propylbenzoimidazol-1-ylmethyl]benzoicAcid (1-a) and4-[5-((R)-1-Benzyl-2-hydroxycarbamoylethylcarbamoyl)-7-methyl-2-propylbenzoimidazol-1-ylmethyl]benzoicAcid (1-b)

To a nitrogen-saturated solution of a mixture of4-[6-((R)-1-benzyl-2-benzyloxycarbamoylethylcarbamoyl)-4-methyl-2-propylbenzoimidazol-1-ylmethyl]benzoicacid and4-[5-((R)-1-benzyl-2-benzyloxycarbamoylethylcarbamoyl)-7-methyl-2-propylbenzoimidazol-1-ylmethyl]benzoicacid, was added 10% Pd/C (200 mg). The mixture was degassed and stirredunder hydrogen (1 atm) overnight at room temperature. The mixture wasfiltered through Celite®, concentrated to dryness, and purified bypreparative reversed phase HPLC. The desired product, compound 1-b, wasisolated as colorless solid (TFA salt; 30 mg).

Compound 1-a: ESMS [M+H]⁺ calcd for C₃₀H₃₂N₄O₅, 529.25; found 529.2.Retention time (anal. HPLC: 10-70% MeCN/H₂O over 5 min)=2.26 min.

Compound 1-b: ESMS [M+H]⁺ calcd for C₃₀H₃₂N₄O₅, 529.25; found 529.2.Retention time (anal. HPLC: 10-70% MeCN/H₂O over 5 min)=2.36 min.

Preparation 4 5-(4′-Bromomethylbiphenyl-2-yl)-1-trityl-1H-tetrazole

To a nitrogen-saturated suspension ofN-triphenylmethyl-5-[4′-methylbiphenyl-2-yl]tetrazole (10 g, 20.9 mmol)in DCM was added NBS (3.7 g, 20.9 mmol) and a catalytic amount ofbenzoyl peroxide (60 mg, 240 μmol). The mixture was stirred at refluxfor 15 hours. After cooling to room temperature, the precipitate wasfiltered and the organic solution was concentrated in vacuo. Silica gelchromatography (EtOAc/hexane) gave the title compound as a white solid.

¹H-NMR (400 MHz, DMSO-d₆): δ (ppm) 4.61 (s, 2H), 6.80 (d, 6H), 7.01 (d,2H), 7.24 (d, 2H), 7.28-7.35 (m, 9H), 7.43-7.45 (dd, 1H), 7.50-7.56 (td,1H), 7.58-7.60 (td, 1H), 7.77-7.79 (dd, 1H).

Preparation 57-Methyl-2-propyl-3-[2′-(1H-tetrazol-5-yl)biphenyl-4-ylmethyl]-3H-benzoimidazole-5-carboxylicacid ((R)-1-benzyl-2-benzyloxycarbamoylethyl)amide (5a) and7-Methyl-2-propyl-1-[2′-(1H-tetrazol-5-yl)biphenyl-4-ylmethyl]-1H-benzoimidazole-5-carboxylicacid ((R)-1-benzyl-2-benzyloxycarbamoylethyl)amide (5b)

To a cold solution of 7-methyl-2-propyl-3H-benzoimidazole-5-carboxylicacid ((R)-1-benzyl-2-benzyloxycarbamoylethyl)amide (800 mg, 825 μmol) inDMF (50 mL) in ice bath was added NaH (60% dispersion in oil; 99 mg, 2.5mmol) in small portions. After stirring for 30 minutes at the sametemperature, 5-(4′-bromomethylbiphenyl-2-yl)-1-trityl-1H-tetrazole (460mg, 825 μmol) was added to the mixture, and the final mixture wasstirred at room temperature for 12 hours, then at 70° C. for 6 hours.The mixture was concentrated in vacuo. The residue was dissolved inEtOAc (200 mL) and washed with saturated aqueous NaCl. The organic layerwas dried over MgSO₄, and evaporated in vacuo, affording a pale yellowoil. The oil was dissolved in DCM (10 mL), followed by the addition ofTFA (10 mL). The final mixture was stirred at room temperature for 1hour, and concentrated to dryness, yielding a pale yellow oil. The oilwas rinsed with ether and dried. The crude material was found to containtwo regioisomeric N-alkylation products, compound 5a (a major product)and compound 5b (a minor desired product). ESMS [M+H]⁺ calcd forC₄₃H₄₂N₈O₃, 719.35; found 719.3.

Example 27-Methyl-2-propyl-3-[2′-(1H-tetrazol-5-yl)biphenyl-4-ylmethyl]-3H-benzoimidazole-5-carboxylicacid ((R)-1-benzyl-2-hydroxycarbamoylethyl)amide (2-a) and7-Methyl-2-propyl-1-[2′-(1H-tetrazol-5-yl)biphenyl-4-ylmethyl]-1H-benzoimidazole-5-carboxylicacid ((R)-1-benzyl-2-hydroxycarbamoylethyl)amide (2-b)

A mixture of7-methyl-2-propyl-3-[2′-(1H-tetrazol-5-yl)biphenyl-4-ylmethyl]-3H-benzoimidazole-5-carboxylicacid ((R)-1-benzyl-2-benzyloxycarbamoylethyl)amide and7-methyl-2-propyl-1-[2′-(1H-tetrazol-5-yl)biphenyl-4-ylmethyl]-1H-benzoimidazole-5-carboxylicacid ((R)-1-benzyl-2-benzyloxycarbamoylethyl)amide were dissolved inEtOH (50 mL), followed by the addition of 10% Pd/C (200 mg). The finalmixture was bubbled with nitrogen gas for 5 minutes, and degassed. Thereaction mixture was stirred under hydrogen (1 atm) for 12 hours, andfiltered through Celite®. The filtrate was concentrated to afford a palebrown oil. The oil was dissolved in 50% aqueous acetic acid, andpurified by reversed phase preparative HPLC. The desired product,compound 2-b (minor component), was isolated as a colorless solid.

Compound 2-a: ESMS [M+H]⁺ calcd for C₃₆H₃₆N₈O₃, 629.30; found 629.4.Retention time (anal. HPLC: 10-70% MeCN/H₂O over 5 min)=2.66 min.

Compound 2-b: ESMS [M+H]⁺ calcd for C₃₆H₃₆N₈O₃, 629.30; found 629.4.Retention time (anal. HPLC: 10-70% MeCN/H₂O over 5 min)=2.75 min.

Preparation 62-Ethoxy-3-[2′-(1H-tetrazol-5-yl)-biphenyl-4-ylmethyl]-3H-benzoimidazole-4-carboxylicacid (1-chloromethyl-3-methylbutyl)amide

1-Chloromethyl-3-methylbutylamine hydrochloride (39 mg, 230 μmol) andtriethylamine (31.6 μL, 1 equiv) were dissolved in DCM (2.00 mL).2-Ethoxy-3-[2′-(1H-tetrazol-5-yl)-biphenyl-4-ylmethyl]-3H-benzimidazole-4-carboxylicacid (100 mg, 1 equiv) was added, followed by HOBt (31 mg, 1 equiv) andEDC HCl (51 mg, 1.1 equiv). The reaction was stirred overnight at roomtemperature, then diluted with DCM (10 mL), washed (H₂O, 10 mL), driedover NaSO₄, decanted, and the solvent evaporated. The residue waspurified by flash chromatography (5% MeOH/DCM) to afford the titlecompound as a white solid (19 mg, 15%). ESMS [M+H]⁺ calcd forC₃₀H₃₂ClN₇O₂ 557.3; found 558.3.

Preparation 7

Thioacetic AcidS-[2-({2-ethoxy-3-[2′-(1H-tetrazol-5-yl)biphenyl-4-ylmethyl]-3H-benzoimidazole-4-carbonyl}amino)-4-methylpentyl]ester

2-Ethoxy-3-[2′-(1H-tetrazol-5-yl)-biphenyl-4-ylmethyl]-3H-benzoimidazole-4-carboxylicacid (1-chloromethyl-3-methylbutyl)amide (100 mg, 180 μmol) andpotassiumthioacetate (31 mg, 15 equiv) were dissolved in MeCN (5 mL) and heatedat 90° C. overnight. The solvent was evaporated and the residuepartitioned between water (10 mL) and EtOAc (10 mL). The aqueous phasewas discarded, the organic layer washed with saturated aqueous NaCl,dried over NaSO₄, decanted, and the solvent evaporated. The crudeproduct (80 mg, 70%) was used without further purification. ESMS [M+H]⁺calcd for C₃₂H₃₅N₇O₃S, 597.3; found 598.3.

Example 32-Ethoxy-3-[2′-(1H-tetrazol-5-yl)-biphenyl-4-ylmethyl]-3H-benzoimidazole-4-carboxylicacid (1-mercaptomethyl-3-methylbutyl)amide

Thioacetic acidS-[2-({2-ethoxy-3-[2′-(1H-tetrazol-5-yl)biphenyl-4-ylmethyl]-3H-benzoimidazole-4-carbonyl}amino)-4-methylpentyl]ester(80 mg, 0.1 mmol) was dissolved in MeOH (5 mL) and 1M NaOH (5 mL). Themixture was stirred for 1 hour with nitrogen gas bubbling through thesolution. The reaction was quenched with acetic acid (5 mL) and themixture evaporated to dryness. The residue was purified using reversephase preparative HPLC to afford the title compound (540 μg). ESMS[M+H]⁺ calcd for C₃₀H₃₃N₇O₂S, 555.3; found 556.4.

Example 4

Following the procedures described in Examples above, and substitutingthe appropriate starting materials and reagents, compounds 4-1 to 4-4,having the following formula, were also prepared:

Ex. R^(2′) R³ R⁶ 4-1 H —(CH₂)₂—CH₃ benzyl 4-2 H —(CH₂)₂—CH₃—CH₂—CH(CH₃)₂ 4-3 H —O—CH₂CH₃ benzyl 4-4 —O—CH₃ —(CH₂)₂—CH₃ benzyl

-   (4-1)    4′-[5-((R)-1-benzyl-2-mercaptoethylcarbamoyl)-2-propylbenzoimidazol-1-ylmethyl]biphenyl-2-carboxylic    acid. MS m/z: [M+H]⁺ calcd for C₃₄H₃₃N₃O₃S, 564.22; found 564.6.-   (4-2)    4′-[5-((R)-1-mercaptomethyl-3-methylbutylcarbamoyl)-2-propylbenzoimidazol-1-ylmethyl]biphenyl-2-carboxylic    acid. MS m/z: [M+H]⁺ calcd for C₃₁H₃₅N₃O₃S, 530.24; found 530.6.-   (4-3)    4′-[5-((R)-1-benzyl-2-mercaptoethylcarbamoyl)-2-ethoxybenzoimidazol-1-ylmethyl]biphenyl-2-carboxylic    acid. MS m/z: [M+H]⁺ calcd for C₃₃H₃₁N₃O₄S, 566.20; found 566.4.-   (4-4)    4′-[5-((R)-1-benzyl-2-mercaptoethylcarbamoyl)-6-methoxy-2-propylbenzoimidazol-1-ylmethyl]biphenyl-2-carboxylic    acid. MS m/z: [M+H]⁺ calcd for C₃₅H₃₅N₃O₄S, 594.24; found 594.4.

Assay 1 AT₁ and AT₂ Radioligand Binding Assays

These in vitro assays were used to assess the ability of test compoundsto bind to the AT₁ and the AT₂ receptors.

Membrane Preparation from Cells Expressing Human AT₁ or AT₂ Receptors

Chinese hamster ovary (CHO-K1) derived cell lines stably expressing thecloned human AT₁ or AT₂ receptors, respectively, were grown in HAM's-F12medium supplemented with 10% fetal bovine serum, 10 μg/mlpenicillin/streptomycin, and 500 μg/ml geneticin in a 5% CO₂ humidifiedincubator at 37° C. AT₂ receptor expressing cells were grown in theadditional presence of 100 nM PD123,319 (AT₂ antagonist). When culturesreached 80-95% confluence, the cells were washed thoroughly in PBS andlifted with 5 mM EDTA. Cells were pelleted by centrifugation and snapfrozen in MeOH-dry ice and stored at −80° C. until further use.

For membrane preparation, cell pellets were resuspended in lysis buffer(25 mM Tris/HCl pH 7.5 at 4° C., 1 mM EDTA, and one tablet of CompleteProtease Inhibitor Cocktail Tablets with 2 mM EDTA per 50 mL buffer(Roche cat.# 1697498, Roche Molecular Biochemicals, Indianapolis, Ind.))and homogenized using a tight-fitting Dounce glass homogenizer (10strokes) on ice. The homogenate was centrifuged at 1000×g, thesupernatant was collected and centrifuged at 20,000×g. The final pelletwas resuspended in membrane buffer (75 mM Tris/HCl pH 7.5, 12.5 mMMgCl₂, 0.3 mM EDTA, 1 mM 5 EGTA, 250 mM sucrose at 4° C.) andhomogenized by extrusion through a 20G gauge needle. Proteinconcentration of the membrane suspension was determined by the methoddescribed in Bradford (1976) Anal Biochem. 72:248-54. Membranes weresnap frozen in MeOH-dry ice and stored at −80° C. until further use.

Ligand Binding Assay to Determine Compound Affinities for the Human AT₁and AT₂ Angiotensin Receptors

Binding assays were performed in 96-well Acrowell filter plates (PallInc., cat.# 5020) in a total assay volume of 100 μL with 0.2 μg membraneprotein for membranes containing the human AT₁ receptor, or 2 μgmembrane protein for membranes containing the human AT₂ receptor inassay buffer (50 mM Tris/HCl pH 7.5 at 20° C., 5 mM MgCl₂, 25 μM EDTA,0.025% BSA). Saturation binding studies for determination of K_(d)values of the ligand were done using N-terminally Europium-labeledangiotensin-TI ([Eu]AngII,H-(Eu—N¹)-Ahx-Asp-Arg-Val-Tyr-Ile-His-Pro-Phe-OH; PerkinElmer, Boston,Mass.) at 8 different concentrations ranging from 0.1 nM to 30 nM.Displacement assays for determination of pK_(i) values of test compoundswere done with [Eu]AngII at 2 nM and 11 different concentrations of drugranging from 1 pM to 10 μM. Drugs were dissolved to a concentration of 1mM in DMSO and from there serially diluted into assay buffer.Non-specific binding was determined in the presence of 10 μM unlabeledangiotensin-II. Assays were incubated for 120 minutes in the dark, atroom temperature or 37° C., and binding reactions were terminated byrapid filtration through the Acrowell filter plates followed by threewashes with 200 μL ice cold wash buffer (50 mM Tris/HCl pH 7.5 at 4° C.,5 mM MgCl₂) using a Waters filtration manifold. Plates were tapped dryand incubated with 50 μl DELFIA Enhancement Solution (PerkinElmer cat.#4001-0010) at room temperature for 5 minutes on a shaker. Filter-bound[Eu]AngII was quantitated immediately on a Fusion plate reader(PerkinElmer) using Time Resolved Fluorescence (TRF). Binding data wereanalyzed by nonlinear regression analysis with the GraphPad PrismSoftware package (GraphPad Software, Inc., San Diego, Calif.) using the3-parameter model for one-site competition. The BOTTOM (curve minimum)was fixed to the value for nonspecific binding, as determined in thepresence of 10 μM angiotensin II. K_(i) values for drugs were calculatedfrom observed IC₅₀ values and the K_(d) value of [Eu]AngII according tothe Cheng-Prusoff equation described in Cheng et al. (1973) BiochemPharmacol. 22(23):3099-108. Selectivities of test compounds for the AT₁receptor over the AT₂ receptor were calculated as the ratio ofAT₂K_(i)/AT₁K_(i). Binding affinities of test compounds were expressedas negative decadic logarithms of the K_(i) values (pK_(i)).

In this assay, a higher pK_(i) value indicates that the test compoundhas a higher binding affinity for the receptor tested. Exemplarycompounds of the invention that were tested in this assay, typicallywere found to have a pK_(i) at the AT₁ receptor greater than or equal toabout 5.0.

Assay 2 In Vitro Assays for the Quantitation of Inhibitor Potencies(IC₅₀) at Human and Rat NEP, and Human ACE

The inhibitory activities of compounds at human and rat NEP and humanACE were determined using in vitro assays as described below.

Extraction of NEP Activity from Rat Kidneys

Rat NEP was prepared from the kidneys of adult Sprague Dawley rats.Whole kidneys were washed in cold PBS and brought up in ice-cold lysisbuffer (1% Triton X-114, 150 mM NaCl, 50 mM Tris pH 7.5; Bordier(1981)J. Biol. Chem. 256:1604-1607) in a ratio of 5 mL of buffer for everygram of kidney. Samples were homogenized using a polytron hand heldtissue grinder on ice. Homogenates were centrifuged at 1000×g in aswinging bucket rotor for 5 minutes at 3° C. The pellet was resuspendedin 20 mL of ice cold lysis buffer and incubated on ice for 30 minutes.Samples (15-20 mL) were then layered onto 25 mL of ice-cold cushionbuffer (6% w/v sucrose, 50 mM pH 7.5 Tris, 150 mM NaCl, 0.06%, TritonX-114), heated to 37° C. for 3-5 minutes and centrifuged at 1000×g in aswinging bucket rotor at room temperature for 3 minutes. The two upperlayers were aspirated off, leaving a viscous oily precipitate containingthe enriched membrane fraction. Glycerol was added to a concentration of50% and samples were stored at −20° C. Protein concentrations werequantitated using a BCA detection system with BSA as a standard.

Enzyme Inhibition Assays

Recombinant human NEP and recombinant human ACE were obtainedcommercially (R&D Systems, Minneapolis, Minn., catalog numbers 1182-ZNand 929-ZN, respectively). The fluorogenic peptide substrate Mca-BK2(Mca-Arg-Pro-Pro-Gly-Phe-Ser-Ala-Phe-Lys(Dnp)-OH; Johnson et al. (2000)Anal. Biochem. 286: 112-118) was used for the human NEP and ACE assays,and Mca-RRL (Mca-DArg-Arg-Leu-(Dnp)-OH; Medeiros et al. (1997) Braz. J.Med. Biol. Res. 30:1157-1162) was used for the rat NEP assay (both fromAnaspec, San Jose, Calif.).

The assays were performed in 384-well white opaque plates at roomtemperature using the respective fluorogenic peptides at a concentrationof 10 μM in assay buffer (50 mM Tris/HCl at 25° C., 100 mM NaCl, 0.01%Tween-20, 1 μM Zn, 0.025% BSA). Human NEP and human ACE were used atconcentrations that resulted in quantitative proteolysis of 5 μM ofMca-BK2 within 20 minutes at room temperature. The rat NEP enzymepreparation was used at a concentration that yielded quantitativeproteolysis of 3 μM of Mca-RRL within 20 minutes at room temperature.

Assays were started by adding 25 μL of enzyme to 12.5 μL of testcompound at 12 concentrations (10 μM to 20 pM). Inhibitors were allowedto equilibrate with the enzyme for 10 minutes before 12.5 μL of thefluorogenic substrates were added to initiate the reaction. Reactionswere terminated by the addition of 10 μL of 3.6% glacial acetic acidafter 20 minutes of incubation. Plates were read on a fluorometer withexcitation and emission wavelengths set to 320 nm and 405 nm,respectively.

Raw data (relative fluorescence units) were normalized to % activityfrom the average high readings (no inhibition, 100% enzyme activity) andaverage low readings (full inhibition, highest inhibitor concentration,0% enzyme activity) using three standard NEP and ACE inhibitors,respectively. Nonlinear regression of the normalized data was performedusing a one site competition model (GraphPad Software, Inc., San Diego,Calif.). Data were reported as pIC₅₀ values.

Exemplary compounds of the invention that were tested in this assay,typically were found to have a pIC₅₀ for the NEP enzyme greater than orequal to about 5.0.

Assay 3 Pharmacodynamic (PD) Assay for ACE, AT₁, and NEP Activity inAnesthetized Rats

Male, Sprague Dawley, normotensive rats are anesthetized with 120 mg/kg(i.p.) of inactin. Once anesthetized, the jugular vein, carotid artery(PE 50 tubing) and bladder (URI-1 urinary silicone catheter) arecannulated and a tracheotomy is performed (Teflon Needle, size 14 gauge)to faciliate spontaneous respiration. The animals are then allowed a 60minute stablization period and kept continuously infused with 5 mL/kg/hof saline (0.9%) throughout, to keep them hydrated and ensure urineproduction. Body temperature is maintained throughout the experiment byuse of a heating pad. At the end of the 60 minute stabilization period,the animals are dosed intravenously (i.v.) with two doses of angiotensin(AngI, 1.0 μg/kg, for ACE inhibitor activity; AngII, 0.1 μg/kg, for AT₁receptor antagonist activity) at 15 minutes apart. At 15 minutespost-second dose of angiotensin (AngI or AngII), the animals are treatedwith vehicle or test compound. Five minutes later, the animals areadditionally treated with a bolus i.v. injection of atrial natriureticpeptide (ANP; 30 μg/kg). Urine collection (into pre-weighted eppendorftubes) is started immediately after the ANP treatment and continued for60 minutes. At 30 and 60 minutes into urine collection, the animals arere-challenged with angiotensin (AngI or AngII). Blood pressuremeasurements are done using the Notocord system (Kalamazoo, Mich.).Urine samples are frozen at −20° C. until used for the cGMP assay. UrinecGMP concentrations are determined by Enzyme Immuno Assay using acommercial kit (Assay Designs, Ann Arbor, Mich., Cat. No. 901-013).Urine volume is determined gravimetrically. Urinary cGMP output iscalculated as the product of urine output and urine cGMP concentration.ACE inhibition or AT₁ antagonism is assessed by quantifying the %inhibition of pressor response to AngI or AngIT, respectively. NEPinhibition is assessed by quantifying the potentiation of ANP-inducedelevation in urinary cGMP output.

Assay 4

In Vivo Evaluation of Antihypertensive Effects in the Conscious SHRModel of Hypertension

Spontaneously hypertensive rats (SHR, 14-20 weeks of age) are allowed aminimum of 48 hours acclimation upon arrival at the testing site. Sevendays prior to testing, the animals are either placed on a restrictedlow-salt diet with food containing 0.11% of sodium for sodium depletedSHRs (SD-SHR) or are placed on a normal diet for sodium repleted SHRs(SR-SHR). Two days prior to testing, the animals are surgicallyimplemented with catheters into a carotid artery and the jugular vein(PE50 polyethylene tubing) connected via a PE10 polyethylene tubing to aselected silicone tubing (size 0.020 ID×0.037 OD×0.008 wall) for bloodpressure measurement and test compound delivery, respectively. Theanimals are allowed to recover with appropriate post operative care.

On the day of the experiment, the animals are placed in their cages andthe catheters are connected via a swivel to a calibrated pressuretransducer. After 1 hour of acclimation, a baseline measurement is takenover a period of at least five minutes. The animals are then dosed i.v.with vehicle or test compound in ascending cumulative doses every 60minutes followed by a 0.3 mL saline to clear the catheter after eachdose. Data is recorded continuously for the duration of the study usingNotocord software (Kalamazoo, Mich.) and stored as electronic digitalsignals. In some studies, the effects of a single intravenous or oral(gavage) dose are monitored for at least 6 hours after dosing.Parameters measured are blood pressure (systolic, diastolic and meanarterial pressure) and heart rate.

Assay 5 In Vivo Evaluation of Antihypertensive Effects in the ConsciousDOCA-Salt Rat Model of Hypertension

CD rats (male, adult, 200-300 grams, Charles River Laboratory, USA) areallowed a minimum of 48 hours acclimation upon arrival at the testingsite before they are placed on a high salt diet.

One week after the start of the high salt diet, a DOCA-salt pellet (100mg, 21 days release time, Innovative Research of America, Sarasota,Fla.) is implanted subcutaneously and unilateral nephrectomy isperformed. On 16 or 17 days post DOCA-salt pellet implantation, animalsare implanted surgically with catheters into a carotid artery and thejugular vein with a PE50 polyethylene tubing, which in turn wasconnected via a PE10 polyethylene tubing to a selected silicone tubing(size 0.020 ID×0.037 OD×0.008 wall) for blood pressure measurement andtest compound delivery, respectively. The animals are allowed to recoverwith appropriate post operative care.

On the day of the experiment, each animal is kept in its cage andconnected via a swivel to a calibrated pressure transducer. After 1 hourof acclimation, a baseline measurement is taken over a period of atleast five minutes. The animals are then dosed i.v. with a vehicle ortest compound in escalating cumulative doses every 60 minutes followedby 0.3 mL of saline to flush the catheter after each dose. In somestudies, the effects of a single intravenous or oral (gavage) dose istested and monitored for at least 6 hours after dosing. Data is recordedcontinuously for the duration of the study using Notocord software(Kalamazoo, Mich.) and stored as electronic digital signals. Parametersmeasured are blood pressure (systolic, diastolic and mean arterialpressure) and heart rate. For cumulative and single dosing, thepercentage change in mean arterial pressure (MAP, mmHg) or heart rate(HR, bpm) is determined as described for Assay 4.

While the present invention has been described with reference tospecific aspects or embodiments thereof, it will be understood by thoseof ordinary skilled in the art that various changes can be made orequivalents can be substituted without departing from the true spiritand scope of the invention. Additionally, to the extent permitted byapplicable patent statues and regulations, all publications, patents andpatent applications cited herein are hereby incorporated by reference intheir entirety to the same extent as if each document had beenindividually incorporated by reference herein.

What is claimed is:
 1. A compound of formula I:

wherein: r is 0, 1 or 2; Ar is an aryl group selected from:

R¹ is selected from —COOR^(1a), —NHSO₂R^(1b), —SO₂NHR^(1d), —SO₂OH,—C(O)NH—SO₂R^(1c), —P(O)(OH)₂, —CN, —OCH(R^(1e))—COOH, tetrazol-5-yl,

R^(1a) is H, —C₁₋₆alkyl, —C₁₋₃alkylenearyl, —C₁₋₃alkyleneheteroaryl,—C₃₋₇cycloalkyl, —CH(C₁₋₄alkyl)OC(O)R^(1aa), —C₀₋₆alkylenemorpholine,

R^(1aa) is —O—C₁₋₆alkyl, —O—C₃₋₇cycloalkyl, —NR^(1ab)R^(1ac), or—CH(NH₂)CH₂COOCH₃; R^(1ab) and R^(1ac) are independently H, —C₁₋₆alkyl,or benzyl, or are taken together as —(CH₂)₃₋₆—; R^(1b) is R^(1c) or—NHC(O)R^(1c); R^(1c) is —C₁₋₆alkyl, —C₀₋₆alkylene-O—R^(1ca),—C5alkylene-NR^(1cb)R^(1cc), —C₀₋₄alkylenearyl, or—C₀₋₄alkyleneheteroaryl; R^(1ca) is H, —C₁₋₆alkyl, or—C₁₋₆alkylene-O—C₁₋₆alkyl; R^(1cb) and R^(1cc) are independently H or—C₁₋₆alkyl, or are taken together as —(CH₂)₂—O—(CH₂)₂— or—(CH₂)₂—N[C(O)CH₃]-(CH₂)₂—; R^(1d) is H, R^(1c), —C(O)R^(1c), or—C(O)NHR^(1c); R^(1e) is —C₁₋₄alkyl or aryl; n is 0, 1, 2 or 3; each R²is independently selected from halo, —NO₂, —C₁₋₆alkyl, —C₂₋₆alkenyl,—C₃₋₇cycloalkyl, —CN, —C(O)R^(2a), —C₀₋₅alkylene-OR^(2b),—C₀₋₅alkylene-NR^(2c)R^(2d), —C₀₋₃alkylenearyl, and—C₀₋₃alkyleneheteroaryl; R^(2a) is H, —C₁₋₆alkyl, —C₃₋₇cycloalkyl,—OR^(2b), or —NR^(2c)R^(2d); R^(2b) is H, —C₁₋₆alkyl, —C₃₋₇cycloalkyl,or —C₀₋₁alkylenearyl; and R^(2c) and R^(2d) are independently H,—C₁₋₄alkyl, or —C₀₋₁alkylenearyl; R^(2′) is selected from H and R²; R³is selected from —C₁₋₁₀alkyl, —C₂₋₁₀alkenyl, —C₃₋₁₀alkynyl,—C₀₋₃alkylene-C₃₋₇cycloalkyl, —C₂₋₃alkenylene-C₃₋₇cycloalkyl,—C₂₋₃alkynylene-C₃₋₇cycloalkyl,—C₀₋₅alkylene-NR^(3a)C₀₋₅alkylene-R^(3b),—C₀₋₅alkylene-O—C₀₋₅alkylene-R^(3b),—C₁₋₅alkylene-S—C₁₋₅alkylene-R^(3b), and —C₀₋₃alkylenearyl; R^(3a) is H,—C₁₋₆alkyl, —C₃₋₇cycloalkyl, or —C₀₋₃alkylenearyl; and R^(3b) is H,—C₁₋₆alkyl, —C₃₋₇cycloalkyl, —C₂₋₄alkenyl, —C₂₋₄alkynyl, or aryl; X is—C₁₋₂alkylene-, where at least one —CH₂— moiety in the alkylene isreplaced with a —NR^(4a)—C(O)— or —C(O)—NR^(4a)— moiety, where R^(4a) isH, —OH, or —C₁₋₄alkyl; R⁵ is selected from —C₀₋₃alkylene-SR^(5a),—C₀₋₃alkylene-C(O)NR^(5b)R^(5c), —C₀₋₃alkylene-NR^(5b)—C(O)R^(5d),—NH—C₀₋₁alkylene-P(O)(OR^(5e))₂, —C₀₋₃alkylene-P(O)OR^(5e)R^(5f),C₀₋₂alkylene-CHR^(5g)—COOH, —C₀₋₃alkylene-C(O)NR^(5h)—CHR^(5i)—COOH, and—C₀₋₃alkylene-S—SR^(5j); R^(5a) is H or —C(O)—R^(5aa); R^(5aa) is—C₁₋₆alkyl, —C₀₋₆alkylene-C₃₋₇cycloalkyl, —C₀₋₆alkylenearyl,—C₀₋₆alkyleneheteroaryl, —C₀₋₆alkylenemorpholine,—C₀₋₆alkylenepiperazine-CH₃, —CH[N(R^(5ab))₂]-aa where aa is an aminoacid side chain, -2-pyrrolidine, —C₀₋₆alkylene-OR^(5ab),—OC₀₋₆alkylenearyl, —C₁₋₂alkylene-OC(O)—C₁₋₆alkyl,—C₁₋₂alkylene-OC(O)—C₀₋₆alkylenearyl, or —C₁₋₂alkylene-OC(O)—OC₁₋₆alkyl;R^(5ab) is independently H or —C₁₋₆alkyl; R^(5b) is H, —OH,—OC(O)R^(5ba), —CH₂COOH, —O-benzyl, -pyridyl, or —OC(S)NR^(5bb)R^(5bc);R^(5ba) is H, —C₁₋₆alkyl, aryl, —OCH₂-aryl, —CH₂O-aryl, or—NR^(5bb)R^(5bc); R^(5bb) and R^(5bc) are independently H or —C₁₋₄alkyl;R^(5c) is H, —C₁₋₁₆alkyl, or —C(O)—R^(5ca); R^(5ca) is H, —C₁₋₆alkyl,—C₃₋₇cycloalkyl, aryl, or heteroaryl; R^(5d) is H, —C₁₋₄alkyl,—C₀₋₃alkylenearyl, —NR^(5da)R^(5db), —CH₂SH, or —O—C₁₋₆alkyl; R^(5da)and R^(5db) are independently H or —C₁₋₄alkyl; R^(5e) is H, —C₁₋₆alkyl,—C₁₋₃alkylenearyl, —C₁₋₃alkyleneheteroaryl, cycloalkyl,—CH(CH₃)OC(O)R^(5ea),

R^(5ea) is —O—C₁₋₆alkyl, —O—C₃₋₇cycloalkyl, —NR^(5eb)R^(5ec), or—CH(NH₂)CH₂COOCH₃; R^(5eb) and R^(5ec) are independently H, —C₁₋₄alkyl,or —C₁₋₃alkylenearyl, or are taken together as —(CH₂)₃₋₆—; R^(5f) is H,—C₁₋₄alkyl, —C₀₋₃alkylenearyl, —C₁₋₃alkylene-NR^(5fa)R^(5fb), or—C₁₋₃alkylene(aryl)- C₀₋₃alkylene-NR^(5fa)R^(5fb); R^(5fa) and R^(5fb)are independently H or —C₁₋₁₄alkyl; R^(5g) is H, —C₁₋₆alkyl,—C₁₋₃alkylenearyl, or —CH₂—O—(CH₂)₂—OCH₃; R^(5h) is H or —C₁₋₄alkyl; andR^(5i) is H, —C₁₋₄alkyl, or —C₀₋₃alkylenearyl; and R^(5j) is—C₁₋₁₆alkyl, aryl, or —CH₂CH(NH₂)COOH; R⁶ is selected from —C₁₋₆alkyl,—CH₂O(CH₂)₂OCH₃, —C₁₋₆alkylene-O—C₁₋₆alkyl, —C₀₋₃alkylenearyl,—C₀₋₃alkyleneheteroaryl, and —C₀₋₃alkylene-C₃₋₇cycloalkyl; and R⁷ is Hor is taken together with R⁶ to form —C₃₋₈cycloalkyl; wherein each —CH₂—group in —(CH₂)_(r)— is optionally substituted with 1 or 2 substituentsindependently selected from —C₁₋₄alkyl and fluoro; each carbon atom inthe alkylene moiety in X is optionally substituted with one or moreR^(4b) groups and one —CH₂— moiety in X may be replaced with a groupselected from —C₄₋₈cycloalkylene-, —CR^(4d)═CH—, and —CH═CR^(4d)—;R^(4b) is —C₀₋₅alkylene-COOR^(4c), —C₁₋₆alkyl, —C₀₋₁alkylene-CONH₂,—C₁₋₂alkylene-OH, —C₀₋₃alkylene-C₃₋₇cycloalkyl, 1H-indol-3-yl, benzyl,or hydroxybenzyl; R^(4c) is H or —C₁₋₄alkyl; and R^(4d) is—CH₂-thiophene or phenyl; each alkyl and each aryl in R¹, R², R^(2′),R³, R^(4a-4d), and R⁵⁻⁶ is optionally substituted with 1 to 7 fluoroatoms; each ring in Ar and each aryl and heteroaryl in R¹, R², R^(2′),R³, and R⁵⁻⁶ is optionally substituted with 1 to 3 substituentsindependently selected from —OH, —C₁₋₆alkyl, —C₂₋₄alkenyl, —C₂₋₄alkynyl,—CN, halo, —O—C₁₋₆alkyl, —S—C₁₋₆alkyl, —S(O)—C₁₋₆alkyl,—S(O)₂—C₁₋₄alkyl, -phenyl, —NO₂, —NH₂, —NH—C₁₋₆alkyl, and—N(C₁₋₆alkyl)₂, wherein each alkyl, alkenyl and alkynyl is optionallysubstituted with 1 to 5 fluoro atoms; or a pharmaceutically acceptablesalt thereof.
 2. The compound of claim 1, wherein r is
 1. 3. Thecompound of claim 1, wherein Ar is:


4. The compound of claim 1, wherein R¹ is —COOH, —NHSO₂R^(1b),—SO₂NHR^(1d), —SO₂OH, —C(O)NH—SO₂R^(1c), —P(O)(OH)₂, —CN,—O—CH(R^(1e))—COOH, tetrazol-5-yl,


5. The compound of claim 4, wherein R¹ is —COOH or tetrazol-5-yl.
 6. Thecompound of claim 1, wherein R¹ is —COOR^(1a), and R^(1a) is —C₁₋₆alkyl,—C₁₋₃alkylenearyl, —C₁₋₃alkyleneheteroaryl, —C₃₋₇cycloalkyl,—CH(C₁₋₄alkyl)OC(O)R^(1aa), —C₀₋₆alkylenemorpholine,


7. The compound of claim 1, wherein n is 0 or n is 1 and R² is—C₁₋₆alkyl.
 8. The compound of claim 1, wherein R^(2′) is H or—C(O)R^(2a), where R^(2a) is —C₁₋₆alkyl.
 9. The compound of claim 1,wherein R³ is —C₁₋₁₀alkyl or —C₀₋₅alkylene-O—C₁₋₅alkylene-R^(3b), whereR^(3b) is H.
 10. The compound of claim 1, wherein X is —C₁₋₂alkylene-,where one —CH₂— moiety in the alkylene is replaced with a —NR^(4a)—C(O)—or —C(O)—NR^(4a)— moiety, where R^(4a) is selected from H, —OH, and—C₁₋₄alkyl.
 11. The compound of claim 10, wherein X is —C(O)—NH—. 12.The compound of claim 1, wherein R⁵ is —C₀₋₃alkylene-SR^(5a),—C₀₋₃alkylene-C(O)NR^(5b)R^(5c), —C₀₋₃alkylene-NR^(5b)—C(O)R^(5d),—NH—C₀₋₁alkylene-P(O)(OR^(5e))₂, —C₀₋₃alkylene-P(O)OR^(5e)R^(5f),—C₀₋₂alkylene-CHR^(5g)—COOH, or —C₀₋₃alkylene-C(O)NR^(5h)—CHR^(5i)—COOH;where R^(5a) is H, R^(5b) is —OH, R^(5c) is H, R^(5d) is H, and R^(5e)is H.
 13. The compound of claim 1, wherein R⁵ is —C₀₋₃alkylene-SR^(5a),—C₀₋₃alkylene-C(O)NR^(5b)R^(5c), —C₀₋₃alkylene-NR^(5b)—C(O)R^(5d),—NH—C₀₋₁alkylene-P(O)(OR^(5e))₂, —C₀₋₃alkylene-P(O)OR^(5e)R^(5f), or—C₀₋₃alkylene-S—SR^(5j); where R^(5a) is —C(O)—R^(5aa); R^(5b) is H,—OC(O)R^(5ba), —CH₂COOH, —O-benzyl, -pyridyl, or —OC(S)NR^(5bb)R^(5bc);R^(5e) is —C₁₋₆alkyl, —C₁₋₃alkylenearyl, —C₁₋₃alkyleneheteroaryl,—C₃₋₇cycloalkyl, —CH(CH₃)—O—C(O)R^(5ea),


14. The compound of claim 12, wherein R¹ is —COOH, —NHSO₂R^(1b),—SO₂NHR^(1d), —SO₂OH, —C(O)NH—SO₂R^(1c), —P(O)(OH)₂, —CN,—O—CH(R^(1e))—COOH, tetrazol-5-yl,


15. The compound of claim 13, wherein R¹ is —COOR^(1a), and R^(1a) is—C₁₋₆alkyl, —C₁₋₃alkylenearyl, —C₁₋₃alkyleneheteroaryl, —C₃₋₇cycloalkyl,—CH(C₁₋₄alkyl)OC(O)R^(1aa), —C₀₋₆alkylenemorpholine,


16. The compound of claim 12, wherein R¹ is —COOR^(1a), and R^(1a) is—C₁₋₆alkyl, —C₁₋₃alkylenearyl, —C₁₋₃alkyleneheteroaryl, —C₃₋₇cycloalkyl,—CH(C₁₋₄alkyl)OC(O)R^(1aa), —C₀₋₆alkylenemorpholine,


17. The compound of claim 13, wherein R¹ is —COOH, —NHSO₂R^(1b),—SO₂NHR^(1d), —SO₂OH, —C(O)NH—SO₂R^(1c), —P(O)(OH)₂, —CN,—O—CH(R^(1e))—COOH, tetrazol-5-yl,


18. The compound of claim 1, wherein R⁶ is selected from —C₁₋₆alkyl and—C₀₋₃alkylenearyl.
 19. The compound of claim 1, wherein R⁷ is H.
 20. Thecompound of claim 1, wherein Ar is substituted with one fluoro atom. 21.The compound of claim 1, having the formula selected from:


22. The compound of claim 1, having the formula

wherein Ar is an aryl group selected from:

R¹ is selected from —COOH and tetrazol-5-yl; n is 0, or n is 1 and R² is—C₁₋₆alkyl; R^(2′) is selected from H and —C(O)—C₁₋₆alkyl; R³ isselected from —C₁₋₁₀alkyl and C₀₋₅alkylene-O—C₁₋₅alkylene-H; X is—C₁₋₂alkylene-, where one —CH₂— moiety in the alkylene is replaced witha —NR^(4a)—C(O)— or —C(O)—NR^(4a)— moiety, where R^(4a) is selected fromH, —OH, and —C₁₋₄alkyl; R⁵ is selected from —C₀₋₃alkylene-SR^(5a) and—C₀₋₃alkylene-C(O)NR^(5b)R^(5c); R^(5a) is H or —C(O)—C₁₋₆alkyl; R^(5b)is H, —OH, or —OC(O)—C₁₋₆alkyl; R^(5c) is H or —C₁₋₆alkyl; and R⁶ is—C₁₋₆alkyl or —C₀₋₃alkylenearyl.
 23. The compound of claim 22, havingthe formula selected from:


24. A pharmaceutical composition comprising a compound of claim 1 and apharmaceutically acceptable carrier.
 25. The pharmaceutical compositionof claim 24, further comprising a second therapeutic agent selected fromthe group comprising diuretics, β₁ adrenergic receptor blockers, calciumchannel blockers, angiotensin-converting enzyme inhibitors, AT₁ receptorantagonists, neprilysin inhibitors, non-steroidal anti-inflammatoryagents, prostaglandins, anti-lipid agents, anti-diabetic agents,anti-thrombotic agents, renin inhibitors, endothelin receptorantagonists, endothelin converting enzyme inhibitors, aldosteroneantagonists, angiotensin-converting enzyme/neprilysin inhibitors,vasopressin receptor antagonists, and combinations thereof
 26. A processfor preparing a compound of claim 1, comprising deprotecting a compoundselected from:

where Ar* is Ar—R¹*; R¹* is selected from —C(O)O—P², —SO₂O—P⁵,—SO₂NH—P⁶, —P(O)(O—P⁷)₂, —OCH(CH₃)—C(O)O—P², —OCH(aryl)-C(O)O—P², andtetrazol-5-yl-P⁴; R⁵* is selected from —C₀₋₃alkylene-S—P³,—C₀₋₃alkylene-C(O)NH(O—P⁵), —C₀₋₃alkylene-N(O—P⁵)—C(O)R^(5d), —C₀₋₁alkylene-NHC(O)CH₂S—P³, —NH—C₀₋₁ alkylene-P(O)(O—P⁷)₂,—C₀₋₃alkylene-P(O)(O—P⁷)—R^(5f), —C₀₋₂alkylene-CHR^(5g)—C(O)O—P²,—C₀₋₃alkylene-C(O)NR^(5h)—CHR^(5i)—C(O)O—P², and —C₀₋₃alkylene-S—S—P³;P² is a carboxy-protecting group; P³ is a thiol-protecting group; P⁴ isa tetrazole-protecting group; P⁵ is a hydroxyl-protecting group; P⁶ is asulfonamide-protecting group; and P⁷ is a phosphate-protecting group orphosphinate-protecting group; or a salt thereof.
 27. An intermediateuseful in the synthesis of a compound of claim 1, selected from thegroup comprising:

where Ar* is Ar—R¹*; R¹* is selected from —C(O)O—P², —SO₂O—P⁵,—SO₂NH—P⁶, —P(O)(O—P⁷)₂, —OCH(CH₃)—C(O)O—P², —OCH(aryl)-C(O)O—P², andtetrazol-5-yl-P⁴; R⁵* is selected from —C₀₋₃alkylene-S—P³,—C₀₋₃alkylene-C(O)NH(O—P⁵), —C₀₋₃alkylene-N(O—P⁵)—C(O)R^(5d), —C₀₋₁alkylene-NHC(O)CH₂S—P³, —NH—C₀₋₁alkylene-P(O)(O—P⁷)₂,—C₀₋₃alkylene-P(O)(O—P⁷)—R^(5f), —C₀₋₂alkylene-CHR^(5g)C(O)O—P²,—C₀₋₃alkylene-C(O)NR^(5h)—CHR^(5i)—C(O)O—P², and —C₀₋₃alkylene-S—S—P³;P² is a carboxy-protecting group; P³ is a thiol-protecting group; P⁴ isa tetrazole-protecting group; P⁵ is a hydroxyl-protecting group; P⁶ is asulfonamide-protecting group; and P⁷ is a phosphate-protecting group orphosphinate-protecting group; or a salt thereof.
 28. A method fortreating hypertension, comprising administering to a patient atherapeutically effective amount of a compound of claim
 1. 29. A methodfor treating heart failure, comprising administering to a patient atherapeutically effective amount of a compound of claim
 1. 30. A methodof treating a patient suffering from a disease or disorder that istreated by antagonizing the AT₁ receptor and/or inhibiting the NEPenzyme, comprising administering to a patient a therapeuticallyeffective amount of a compound of claim 1.